Composition for preventing or treating tnf-related diseases, containing novel derivative as active ingredient, and method for inhibiting tnf activity by using same

ABSTRACT

The present invention relates to: a 4-benzopyranone derivative and a pharmaceutically acceptable salt, solvate, racemate, or stereoisomer thereof; a composition for preventing, alleviating or treating TNF-related diseases, containing the same as an active ingredient; and a method for treating TNF-related diseases, a reagent composition for inhibiting TNF, and a method for inhibiting TNF, all of which use the same. The compositions can be orally administered so as not to cause injection side effects, do not cause immunological tolerance, and can effectively inhibit a TNF activity by being directly bound to TNF while facilitating co-administering with a conventional oral preparation and dosage control.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Rule 53(b) Continuation of U.S. application Ser.No. 16/757,112 filed Apr. 17, 2020, which is a National Stage ofInternational Application No. PCT/KR2018/007922 filed Jul. 12, 2018,which claims priority based on Korean Patent Application No.10-2017-0135899 filed Oct. 19, 2017, the respective disclosures of whichare incorporated herein by reference in their entirety.

TECHNICAL FIELD

The present invention relates to a novel 4-benzopyranone compoundderivative, and a composition for preventing, alleviating or treatingTNF-related diseases, containing the same as an active ingredient, and amethod for inhibiting a TNF activity by using the same.

BACKGROUND ART

TNF is a key inflammatory cytokine, and is attracting attention as amajor cause of autoimmune inflammatory diseases such as rheumatoidarthritis, inflammatory bowel disease, psoriasis, ankylosing spondylitisand the like. A variety of anti-TNF blockbuster biopharmaceuticals suchas etanercept (Enbrel), adalimumab (Humira), infliximab (Remicade) andthe like are actively used as a therapeutic agent in a variety ofTNF-related diseases including rheumatoid arthritis. Anti-TNFbiopharmaceuticals show an excellent efficacy in a short period of time,but they are expensive and require repetitive injections, so patientsare highly repulsed. In addition, about one third of patients have notherapeutic effect, and tolerance occurs within a few years due toimmunological side effects even in patients who have responded to thedrugs. There are obvious disadvantages such as difficulty of storagebecause it is necessary to store them at low temperature. Anti-TNFbiopharmaceuticals have unmet medical demand.

As a strategy to overcome this, efforts have been made to find smallmolecular materials that can be orally administered and act likeantibodies by being directly bound to cytokines or their receptors [Heet al. (2005). Science 310(5750): 1022-1025]. However, binding tocytokines and their receptors occurs as a bond between protein andprotein in a large area, and inhibiting the same by small molecularmaterials has not been effective so far.

An article regarding to a small molecular material that dissociates theTNF trimer by being directly bound to TNF has been published [He et al.(2005). Science 310(5750): 1022-1025], a follow-up study seems to bestopped due to weak activity and the like. On the one hand, many smallmolecular material signaling inhibitors that block intracellularsignaling are under development, and articles on the inhibitors of TNFexpression or secretion that have unclear mechanisms have beenpublished, but there is no data of in vivo activity. The development ofsmall molecular material drugs that directly inhibit the binding betweenTNF and TNFR has not been successful so far.

DETAILED DESCRIPTION OF THE INVENTION Technical Problem

It is an object of the present invention to provide a novel compoundwhich shows an excellent inhibitory effect on a TNF activity.

It is another object of the present invention to provide apharmaceutical composition which can effectively prevent or treatdiseases by showing an excellent inhibitory effect on a TNF activity.

It is still another object of the present invention to provide a healthfunctional food composition which can effectively prevent or alleviatediseases by showing an excellent inhibitory effect on a TNF activity.

It is still another object of the present invention to provide a methodfor treating TNF-related diseases effectively.

It is still another object of the present invention to provide a reagentcomposition for inhibiting a TNF activity in vitro.

It is still another object of the present invention to provide a methodfor inhibiting a TNF activity in an animal excluding a human.

Solution to Problem

In order to accomplish the object described above, the present inventionprovides a 4-benzopyranone derivative represented by Formula 1 below, apharmaceutically acceptable salt, solvate, racemate, or stereoisomerthereof.

In order to accomplish another object described above, the presentinvention provides a pharmaceutical composition for preventing ortreating TNF-related diseases, wherein the pharmaceutical compositioncontains a 4-benzopyranone derivative represented by Formula 1 below, apharmaceutically acceptable salt, solvate, racemate, or stereoisomerthereof as an active ingredient, and inhibits a TNF activity by beingdirectly bound to tumor necrosis factor (TNF).

In order to accomplish still another object described above, the presentinvention provides a health functional food composition preventing oralleviating TNF-related diseases, wherein the health functional foodcomposition contains a 4-benzopyranone derivative represented by Formula1 below, a pharmaceutically acceptable salt, solvate, racemate, orstereoisomer thereof as an active ingredient, and inhibits a TNFactivity by being directly bound to tumor necrosis factor (TNF).

In order to accomplish still another object described above, the presentinvention provides a method for treating TNF-related diseases,comprising a step of treating with a 4-benzopyranone derivativerepresented by Formula 1 below, a pharmaceutically acceptable salt,solvate, racemate, or stereoisomer thereof in a pharmaceuticallyeffective amount.

In order to accomplish still another object described above, the presentinvention provides a reagent composition for inhibiting a TNF activityin vitro, containing a 4-benzopyranone derivative represented by Formula1 below, a pharmaceutically acceptable salt, solvate, racemate, orstereoisomer thereof as an active ingredient.

In order to accomplish still another object described above, the presentinvention provides a method for inhibiting a TNF activity, comprising astep of treating an animal excluding a human with a 4-benzopyranonederivative represented by Formula 1 below, a pharmaceutically acceptablesalt, solvate, racemate, or stereoisomer thereof.

in which,

R¹ is heteroaryl or substituted phenyl;

R² is hydrogen or lower alkyl of (C1-C4); and

R³, R⁴, R⁵, and R⁶ are each independently hydrogen, halogen, or(C1-C4)alkoxy.

Effect of Invention

The present invention relates to a composition for preventing ortreating TNF-related diseases, containing a 4-benzopyranone derivativecompound as an active ingredient. The conventional proteinTNF-inhibiting biopharmaceuticals are inconvenient for theadministration in combination with other compound therapeutic agents,and can not be utilized for the development of a combination product.However, the TNF-inhibiting compound provided by the present inventionis very easy to be administered in combination with other conventionalcompound therapeutic agents or to be utilized for the development of acombination product, and, in addition, has advantages such as excellentefficacy, low cost, non-invasive oral administration,non-immunogenicity, needlessness of refrigeration storage and the like,and thus, can be usefully utilized in a composition for preventing ortreating TNF-related diseases.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 is a graph showing the TNF-inhibiting ability of representativecompounds inhibiting TNF (the compound of Example 46, the compound ofExample 61, and the compound of Example 51) among the small molecularcompounds synthesized in one example of the present invention dependingon concentration.

FIGS. 2 a to 2 e show the competitive inhibitory activity against TNFdirect binding and TNF-etanercept binding of the compounds of thepresent invention ((FIG. 2 a ): the results of confirming the binding ofthe compound of Example 61 with TNF bound to chips by the surfaceplasmon resonance analysis method (Biacore T200), (FIG. 2 b ): theresults of measuring the binding affinity (K_(D)) of the compound ofExample 61 with TNF bound to chips, (FIG. 2 c ): the results ofmeasuring whether the compound binds to a TNF receptor, etanercept bythe surface plasmon resonance analysis method, (FIG. 2 d ) the resultsof measuring the binding affinity (K_(D)) of the compound of Example 30for TNF which has a weak TNF-inhibiting efficacy as a control group, and(FIG. 2 e ): the results of confirming the inhibiting ability of thecompound of Example 61 on the TNF-etanercept binding by the surfaceplasmon resonance analysis method).

FIGS. 3 a and 3 b show (FIG. 3 a ) the inhibitory activity against thebinding between TNF-cell of the TNF-inhibiting compounds, and (FIG. 3 b) the inhibitory activity against a TNF-induced signaling of thecompound of Example 61 according to one example of the presentinvention.

FIGS. 4 a and 4 b show the sepsis therapeutic effect of theTNF-inhibiting compound according to one example of the presentinvention ((FIG. 4 a ): the results of measuring the in vivoTNF-inhibitory activity of the compound of Example 61 in a TNF-inducedsepsis lethality model, and (FIG. 4 b ): the results of experimentingthe in vivo efficacy on the administration of a TNF-inhibiting compound,the compound of Example 61 in combination with an IL inhibitingcompound, LMT-28 using the lipopolysaccharide (LPS)-induced sepsismodel).

FIGS. 5 a to 5 c shows the effect of preventing or treating rheumatoidarthritis of the TNF-inhibiting compound according to one example of thepresent invention ((FIG. 5 a ): the results of confirming the effect ofpreventing rheumatoid arthritis of the compound of Example 61 in aTNF-overexpressing mouse model, (FIG. 5 b ): the results of confirmingthe effect of treating rheumatoid arthritis of the compound of Example61 in a TNF-overexpressing mouse model, and (FIG. 5 c ): the results ofconfirming the effect of treating rheumatoid arthritis of the compoundof Example 61 in a CIA mouse model).

FIG. 6 shows the therapeutic effect when co-administering the compoundsin an animal model of dextran sulfate sodium (DSS) induced inflammatorybowel disease.

FIG. 7 shows the effect of protecting acute kidney injury of thecompounds in an animal model of cecal ligation and puncture(CLP)-induced sepsis.

FIG. 8 shows the results of confirming the synergistic therapeuticeffect when co-administering the compound of Example 61 and methotrexate(MTX) in an animal model of rheumatoid arthritis.

BEST EMBODIMENT FOR WORKING THE INVENTION

Hereinafter, the present invention will be described in more detail.

The present invention provides a 4-benzopyranone derivativecharacterized by being represented by Formula 1, a pharmaceuticallyacceptable salt, solvate, racemate, or stereoisomer thereof:

in which,

R¹ is heteroaryl or substituted phenyl;

R² is hydrogen or lower alkyl of (C1-C4); and

R³, R⁴, R⁵, and R⁶ are each independently hydrogen, halogen, or(C1-C4)alkoxy.

In one example of the present invention, a 4-benzopyranone derivativehaving a carboxy group at the 3-position as described above among the4-benzopyranone compounds can be prepared through the method disclosedin Reaction Scheme 1 below.

A step of preparing the compound (3) by reacting the compound (2) withan acid chloride in the presence of a base and a catalyst (Step 1); astep of preparing the compound (4) by reacting the compound (3) preparedin Step 1 above through the intermolecular Wittig reaction (Step 2); anda step of preparing the compound (5) by de-esterifying the compound (4)prepared in Step 2 above under an acidic condition to remove thetertiary butyl group (Step 3).

Each preparation step of the present invention will be describedspecifically as follows: Step 1 is a step of preparing the compound (3)by reacting the compound (2) with heteroaromatic or benzoyl chloride inthe presence of a base and a catalyst.

N,N-diisopropylethylamine, triethylamine, or pyridine may be used as thebase, and 4-dimethylaminopyridine may be used as the catalyst. The acidchloride is used in a slight excess amount, preferably 1.05 equivalents,to result in the esterification reaction. It is possible to remove theacid chloride remaining after the reaction by the purification usingchromatography or the reaction work-up method using a Girard-T(H₂NNHCOCH₂N(CH₃)₃Cl) reagent instead of chromatography. The acidchloride may be removed easily because the acid chloride reacts with theabove Girard-T reagent to form a hydrazine derivative which is dissolvedin water, and only the desired compound (3) remains in the organiclayer, and all of the remains are contained in the aqueous layer.

The compound (2), which is the starting material of Step 1, may beobtained by reacting an O-acetylsalicylic acid chloride compound with atertiary butoxy carbonyl methylene triphenylphosphorane compound in thepresence of a reaction solvent of benzene or tetrahydrofuran and acatalyst of N,O-bistrimethylsilyl acetamide to prepare a phosphoranecompound, and then, by removing an acetyl group through a deacetylationreaction using a base of dimethylamine, methylamine, or ammonia and areaction solvent of tetrahydrofuran, methanol, or ethanol.

Next, Step 2 is a step of preparing the compound (4) by reacting thecompound (3) prepared in Step 1 above through the intermolecular Wittigreaction. In this regard, benzene, toluene, xylene, or mesitylene may beused as a reaction solvent, and it is preferable to reflux with heatingat 80 to 140° C. for 8 to 24 hours.

Next, Step 3 is a step of preparing the compound (5) by de-esterifyingthe compound (4) prepared in Step 2 above under an acidic condition toremove the tertiary butyl group. The de-esterification reaction may becarried out by reacting the compound with trifluoracetic acid in acarbon dichloride solvent. By the reaction, the tertiary butyl group maybe removed from the carboxylic acid butyl ester compound (4) to obtainthe 4-benzopyranone compound (5) having a carboxy group at the3-position in Formula 1.

In addition to the compounds specified as the base material, thesolvent, and the catalyst in each of the above-mentioned reactions, acompound capable of promoting the reaction may be used as a basematerial, a solvent, and a catalyst.

The products after the completion of each reaction or all reactions asdescribed above may be separated and purified by a customary work-upmethod, for example, chromatography or recrystallization.

The compound of Formula 1 of the present invention can be synthesized byusing the synthesis methods described above and can be obtained by allcustomary methods, and commercially available reagents can be also used.

The 4-benzopyranone compound of the present invention may be in the formof a pharmaceutically acceptable salt thereof The salt includes thecustomary acid addition salts used in the field of compound inhibitors,for example, salts derived from inorganic acids such as hydrochloricacid, bromic acid, sulfuric acid, sulfamic acid, phosphoric acid, ornitric acid, and salts derived from organic acids such as acetic acid,propionic acid, succinic acid, glycolic acid, stearic acid, citric acid,maleic acid, malonic acid, methanesulfonic acid, tartaric acid, malicacid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid,2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, oxalic acid,or trifluoroacetic acid. In addition, the salt includes salts derivedfrom metals such as, for example, lithium, sodium, potassium, magnesium,or calcium in the form of customary metal salts. The acid addition saltsor metal salts may be prepared according to customary methods known inthe field of organic chemistry.

In addition, the 4-benzopyranone compound of the present invention maybe in the form of a solvate thereof The term “solvate” means a complexor aggregate formed by one or more solute molecules, i.e., a compound ofFormula 1 or a pharmaceutically acceptable salt thereof, and one or moresolvent molecules. The solvate may be, for example, a complex oraggregate formed with water, methanol, ethanol, isopropanol, or aceticacid.

In addition, the 4-benzopyranone compound of the present invention maybe in the form of a stereoisomer thereof. The stereoisomer includes allstereoisomers such as enantiomers and diastereomers. The compound may bea stereoisomerically pure form or a mixture of one or morestereoisomers, for example, a racemic mixture. The separation of certainstereoisomers can be carried out by any one of customary methods knownin the art. Specifically, in Formula 1 above, R¹ may be furanyl,thiophenyl, or substituted phenyl represented by

and R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ may be each independentlyhydrogen, chloro, bromo, fluoro, or methoxy.

More specifically, the 4-benzopyranone derivative may be selected fromthe group consisting of2-(2-chlorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy chloro-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl) carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy chloro-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-bromo-(4H) benzopyranone;2-(4-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone; and2-(4-chlorophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone.

In one example of the present invention, the 4-benzopyranone derivativecompound may inhibit a TNF activity by being directly bound to tumornecrosis factor (TNF).

Furthermore, the present invention provides a pharmaceutical compositionfor preventing or treating TNF-related diseases or a health functionalfood composition for preventing or alleviating TNF-related diseases,wherein the composition contains a 4-benzopyranone derivativerepresented by Formula 1, a pharmaceutically acceptable salt, solvate,racemate, or stereoisomer thereof as an active ingredient, and inhibitsa TNF activity by being directly bound to tumor necrosis factor (TNF):

in which,

R¹ is heteroaryl or substituted phenyl;

R² is hydrogen or lower alkyl of (C1-C4); and

R³, R⁴, R⁵, and R⁶ are each independently hydrogen, halogen, or(C1-C4)alkoxy.

Specifically, in Formula 1 above, R¹ may be furanyl, thiophenyl, orsubstituted phenyl represented by

and R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ may be each independentlyhydrogen, chloro, bromo, fluoro, or methoxy.

More specifically, the 4-benzopyranone derivative may be selected fromthe group consisting of2-(2-chlorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl) carboxy-8-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3,4-dim ethoxyphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3,5-difluorophenyl) carboxy-6-bromo-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl) carboxy-7-chloro-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl) carboxy-7-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone; and2-(4-chlorophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone.

In this regard, the TNF-related disease may be any one selected from thegroup consisting of autoimmune diseases, inflammatory diseases,cardiovascular diseases, metabolic diseases, immune disorders,neurological diseases, ophthalmic diseases, skin diseases, psychiatricdiseases, infectious diseases, and cancers, but is not limited thereto.

Specifically, the TNF-related disease may be any one selected from thegroup consisting of rheumatoid arthritis, juvenile rheumatoid arthritis,inflammatory bowel disease, Crohn's disease, ulcerative colitis,psoriasis, plaque psoriasis, juvenile plaque psoriasis, psoriaticarthritis, polyarticular juvenile idiopathic arthritis, Behcet'senteritis, ankylosing spondylitis, axial spondyloarthritis, juvenileenthesitis-related arthritis, polymyalgia rheumatica, multiplesclerosis, thyroiditis, delayed hypersensitivity, allergy, contactdermatitis, atopic dermatitis, systemic lupus erythematosus, systemicsclerosis, adult-onset Still's disease, asthma, autoimmune thyroiddisorder, Sjogren's syndrome, Kawasaki disease, pancreatitis, nephritis,hepatitis, pneumonia, chronic obstructive pulmonary disease, otitismedia, angioplasia nephritis, myelodysplastic syndrome, osteoarthritis,sarcoidosis, granuloma annulare, Wegener's granulomatosis, lupus,hemolytic uremic syndrome, arteriosclerosis, vasculitis, heart failure,stroke, myocardial infarction, myocardial ischemia-reperfusion injury,sexual dysfunction, obesity, hypertension, diabetes mellitus anddiabetic complication, hyperlipidemia, preeclampsia, kidney disease,liver disease, kidney injury, liver injury, snake bite, allograftrejection, organ transplantation, graft versus host disease, dementia,Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis,pain, central nervous system disease, uveitis, Behcet's disease,diabetic macular edema, macular degeneration, orbitopathy, glaucoma,hidradenitis suppurativa, multicentric reticulohistiocytosis, pityriasisrubra pilaris, eosinophilic fascilitis, panniculitis, necrobiosislipoidica diabeticorum, cicatricial pemphigoid, pyoderma gangrenosum,Sweet's syndrome, subcorneal pustular dermatosis, scleroderma,neutrophilic dermatitis, toxic epidermal necrolysis, pustulardermatitis, dermatomyositis, polymyositis, bullous dermatosis, erythemanodosum, alopecia, depressive disorder, bipolar disorder, anxietydisorder, tuberculosis, viral infection, bacterial infection, fungalinfection, protozoan infection, cerebral malaria, sepsis, septic shock,prostate cancer, skin cancer, colorectal cancer, kidney cancer,pancreatic cancer, ovarian cancer, breast cancer, bladder cancer,prostate cancer, lymphoma, glioma, osteosarcoma, leukemia, multiplemyeloma, and cachexia, but is not limited thereto.

In one example of the present invention, the composition may furthercontain a drug, wherein the administration in combination with the drugcan more effectively prevent, alleviate, or treat TNF-related diseases.

The “drug” used in the present invention is a substance capable ofinducing a desired biological or pharmacological effect by promoting orinhibiting a physiological function in the body of an animal or human,and means a chemical or biological substance or compound suitable to beadministered to an animal or human, and may (1) have a prophylacticeffect on the organism by the prevention of undesired biological effectssuch as the prevention of infection, (2) ameliorate the condition causedby the disease, for example, mitigate the pain or infection resultingfrom the disease, and (3) play a role in mitigating, reducing, orcompletely eliminating the disease from the organism.

Specifically, the drug may be selected from anti-rheumatic drugs(DMARDs), nonsteroidal anti-inflammatory drugs (NSAIDs), steroids,antimetabolites, anti-inflammatory agents, antibiotics, signaling/enzymeinhibitors, receptor inhibitors, HMGB1 inhibitors, antithrombotic drugs,autophagy agonists, cytokine inhibitors, HMG-CoA reductase inhibitors,antihypertensive agents, anticancer agents, immune activation agents, Bcell inhibitors, and T cell inhibitors, but is not limited thereto.

More specifically, the drug may be selected from the group consisting ofmethotrexate, hydroxychloroquine, sulfasalazine, leflunomide,bucillamine, cyclosporine, tacrolimus, azathioprine, cyclophosphamide,mizoribine, penicillamine, oral gold preparation, antimalarial agent,6-mercaptopurine, indomethacin, naproxen, sulindac, diclofenac,aceclofenac, mefenamic acid, aspirin, fenoprofen, salsalate, piroxicam,etodolac, flurbiprofen, ibuprofen, loxoprofen, nabumetone, lonazolac,meloxicam, fenbufen, ketorolac tromethamine, indoprofen, ketoprofen,suprofen, carprofen, tiaprofenic acid, flufenamic acid, ebselen,felbinac, tolmetin, flunixin, celecoxib, rofecoxib, hydrocortisone,cortisone, prednisolone, methylprednisolone, triamcinolone,betamethasone, dexamethasone, fludrocortisone, entocort,5-aminosalicylate, 6-thioguanine, cytarabine, 5-fluorouracil,dacarbazine, mechlorethamine, thiotepa, chlorambucil, melphalan,carmustine, lomustine, busulfan, sestrin 2, withaferin A, celastrol,quercetin, luteolin, curcumin, metformin, dibromomannitol, GR270773,pentoxifylline, N-acetylcysteine, melatonin, resveratrol, mesalamine,single chain fatty acid, glutamine, gemfibrozil, retinoid, hydroxyurea,trihydroxyisoflavone, deoxykaempferol, kaempferol, gingerol, caffeicacid, cyanidin, cryptotanshinone, deguelin, delphinidin, equol, fisetin,myricetin, procyanidin B2, metronidazole, ciprofloxacin, niclosamide,thiabendazole, imipenem-cilastatin, fluoroquinolone, tofacitinib,glyburide, rolipram, doxycycline, VX-166, zVAD, L-97-1, ISO-1,tauroursodeoxycholic acid, HK-156, A-285222, CP-0127,Bis-N-norgliovictin, aurintricarboxylic acid, chloroamidine, ouabain,terazosin, prazosin, tranilast, apremilast, monobenzone,phenazopyridine, 546C88, NOX-100, gabexate mesilate, ulinastatin,somatostatin, octreotide, IKK inhibitor, caspase inhibitor, TAK-242,eritoran, ki16425, camptothecin, caffeic acid phenethyl ester,sulforaphane, Tim-3, BN-52021, BB-882, TCV-309, CT-400, ethyl pyruvate,hemin, CORM-2, tanshinone IIA sulfonate, nicotine, EGCG,isorhamnetin-3-O-galactoside, persicarin, catechin, carbenoxolone,glycyrrhizin, emodin-6-O-b-D-glucoside, acteoside, forsythoside B,rosmarinic acid, chlorogenic acid, inflachromene, cilostazol,clopidogrel, sarpogrelate, drotrecogin alpha, carbamazepine,chloroquine, anakinra, tocilizumab, LMT-28,1-(3-dimethylaminopropyl)-3-ethylurea, gp13 OF c, beta-arrestin 2,IL-30, diacerein, secukinumab, ustekinumab, ixekizumab, thalidomide,adalimumab, infliximab, pravastatin, atorvastatin, rosuvastatin,simvastatin, losartan, telmisartan, hydrochlorothiazide, furosemide,propranolol, metoprolol, captopril, amlodipine, clonidine, methyldopa,minoxidil, streptozotocin, mitomycin, cisplatin, daunorubicin,doxorubicin, dactinomycin, bleomycin, mithramycin, anthramycin,calicheamicin, duocarmycin, vincristine, taxol, docetaxel, cytochalasinB, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide,teniposide vinblastine, colchicine, dihydroxy anthracenedione,mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone,procaine, tetracaine, lidocaine, propranolol, tamoxifen, bazedoxifene,puromycin, anetholtrithion, nivolumab, pembrolizumab, ipilimumab,atezolizumab, alpha-galactosylceramide, SRT3025, DTA-1, IL-7, IL-2,IL-15, CXCL1, ATRA, gemcitabine, carboplatin, NCX-4016, CDDO-Me,sunitinib, zoledronic acid, Astragalus polysaccharide, rituximab,imuran, abatacept, GW9662, rosiglitazone, Y-27632, and alefacept, but isnot limited thereto.

The composition may contain from 0.0001% by weight to 10% by weight,based on the total weight of the composition, of a 4-benzopyranonederivative, a pharmaceutically acceptable salt, solvate, racemate, orstereoisomer thereof, preferably 0.001% by weight to 1% by weight, butnot limited thereto.

In one embodiment of the present invention, the pharmaceuticalcomposition for preventing or treating TNF-related diseases, containingthe 4-benzopyranone derivative, pharmaceutically acceptable salt,solvate, racemate, or stereoisomer thereof as an active ingredient maybe used in the form of any one of the formulations selected from thegroup consisting of injections, granules, powders, tablets, pills,capsules, suppositories, gels, suspensions, emulsions, drops, orsolutions according to a customary method.

In another embodiment of the present invention, the pharmaceuticalcomposition may further contain one or more additives selected from thegroup consisting of carriers, excipients, disintegrating agents,sweetening agents, coating agents, bulking agents, glidants, flavoringagents, antioxidants, buffers, bacteriostatic agents, diluents,dispersing agents, surfactants, binders, and lubricants, which areappropriate to be customarily used in the preparation of thepharmaceutical composition.

Specifically, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol,erythritol, maltitol, starch, acacia gum, alginate, gelatin, calciumphosphate, calcium silicate, cellulose, methyl cellulose,microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, andmineral oil may be used as the carrier, excipient, and diluent. Solidpreparations for oral administration include tablets, pills, powders,granules, capsules and the like. These solid preparations may beprepared by mixing at least one or more excipients, for example, starch,calcium carbonate, sucrose or lactose, gelatin and the like into thecomposition. In addition to simple excipients, lubricants such asmagnesium stearate, talc may be also used. Liquid preparations for oraladministration include suspensions, solutions for internal use,emulsions, syrups and the like, and may comprise various excipients, forexample wetting agents, sweetening agent, perfuming agents,preservatives and the like, in addition to water and liquid paraffinwhich are commonly used simple diluents. Preparations for parenteraladministration include sterilized aqueous solutions, non-aqueoussolutions, suspensions, emulsions, lyophilized preparations,suppositories and the like. As non-aqueous solvents or suspendingagents, propylene glycol, polyethylene glycol, vegetable oil such asolive oil, injectable ester such as ethyl oleate and the like may beused. As the base of the suppositories, witepsol, Macrogol, Tween 61,cacao butter, laurin butter, glycerogelatin and the like may be used.

The preferred dosage of the 4-benzopyranone derivative, pharmaceuticallyacceptable salt, solvate, racemate, or stereoisomer thereof may varydepending on the condition and body weight of the subject, the kind andseverity of the disease, the form of the drug, administration route, andthe duration, and may be appropriately selected by those skilled in theart. However, for the preferred effect, the compound of the presentinvention may be administered in a daily dosage of 0.0001 to 100 mg/kg,preferably 0.001 to 100 mg/kg. The dose may be administered once per dayor in several divided doses per day, and the scope of the presentinvention is not limited thereto.

In one embodiment of the present invention, the pharmaceuticalcomposition may be administered to a mammal such as a rat, a mouse, alivestock, a human and the like in a variety of routes. All modes ofadministration may be expected, and the pharmaceutical composition maybe administered, for example, by oral, rectal or intravenous,intramuscular, subcutaneous, intrauterine, intrathecal, orintracerebroventricular injections.

In one embodiment of the present invention, the health functional foodcomposition may contain various nutrients, vitamins, minerals(electrolytes), flavors such as synthetic flavors and natural flavors,coloring agents and improving agents (cheese, chocolate and the like),pectic acid and salts thereof, alginic acid and salts thereof, organicacids, protective colloidal thickening agents, pH adjusting agents,stabilizing agents, preservatives, glycerin, alcohols, carbonizingagents as used in carbonated beverages and the like. Additionally, thehealth functional food composition may contain fruit flesh for thepreparation of natural fruit juices, synthetic fruit juices, andvegetable beverages. These components may be used independently or incombination. In addition, the health functional food composition may bein the form of any one of meats, sausages, bread, chocolate, candies,snacks, confectionery, pizza, ramen, gum, ice cream, soups, beverages,teas, functional water, drinks, alcoholic beverages, and multi-vitaminpreparations.

In addition, the health functional food composition may further comprisea food additive, and the suitability of the additive as a “foodadditive” is determined based on a standard and criteria relating to theconcerned item according to general rules and general test methods andthe like of the Korean Food Additives Code that has been approved by theMinistry of Food and Drug Safety as long as other rules are notprovided.

The items disclosed in the “Korean Food Additives Code” include, forexample, a chemically synthetic product, such as ketone, glycine,calcium citrate, nicotinic acid, cinnamic acid and the like, a naturaladditive product, such as persimmon color, a licorice extract,microcrystalline cellulose, Kaoliang color, guar gum and the like, andmixed preparations, such as a sodium L-glutamate preparation, an alkaliagent for noodles, a preservative preparation, a tar color preparationand the like.

In this regard, the content of the 4-benzopyranone derivative,pharmaceutically acceptable salt, solvate, racemate, or stereoisomerthereof which is added to foods during the preparation of the healthfunctional food composition may be appropriately increased or decreased,if necessary, and may be added to be included preferably in an amount of1% by weight to 90% by weight.

Furthermore, the present invention provides a method for treatingTNF-related diseases, comprising a step of treating with a4-benzopyranone derivative represented by Formula 1, a pharmaceuticallyacceptable salt, solvate, racemate, or stereoisomer thereof in apharmaceutically effective amount:

in which,

R¹ is heteroaryl or substituted phenyl;

R² is hydrogen or lower alkyl of (C1-C4); and

R³, R⁴, R⁵, and R⁶ are each independently hydrogen, halogen, or(C1-C4)alkoxy.

Specifically, in Formula 1 above, R¹ may be furanyl, thiophenyl, orsubstituted phenyl represented by

and R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ may be each independentlyhydrogen, chloro, bromo, fluoro, or methoxy.

More specifically, the 4-benzopyranone derivative may be selected fromthe group consisting of2-(2-chlorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3,4-dim ethoxyphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3,5-difluorophenyl) carboxy-6-bromo-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl) carboxy-7-chloro-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone; and2-(4-chlorophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone.

In this regard, the TNF-related disease may be any one selected from thegroup consisting of autoimmune diseases, inflammatory diseases,cardiovascular diseases, metabolic diseases, immune disorders,neurological diseases, ophthalmic diseases, skin diseases, psychiatricdiseases, infectious diseases, and cancers, but is not limited thereto.

Specifically, the TNF-related disease may be any one selected from thegroup consisting of rheumatoid arthritis, juvenile rheumatoid arthritis,inflammatory bowel disease, Crohn's disease, ulcerative colitis,psoriasis, plaque psoriasis, juvenile plaque psoriasis, psoriaticarthritis, polyarticular juvenile idiopathic arthritis, Behcet'senteritis, ankylosing spondylitis, axial spondyloarthritis, juvenileenthesitis-related arthritis, polymyalgia rheumatica, multiplesclerosis, thyroiditis, delayed hypersensitivity, allergy, contactdermatitis, atopic dermatitis, systemic lupus erythematosus, systemicsclerosis, adult-onset Still's disease, asthma, autoimmune thyroiddisorder, Sjogren's syndrome, Kawasaki disease, pancreatitis, nephritis,hepatitis, pneumonia, chronic obstructive pulmonary disease, otitismedia, angioplasia nephritis, myelodysplastic syndrome, osteoarthritis,sarcoidosis, granuloma annulare, Wegener's granulomatosis, lupus,hemolytic uremic syndrome, arteriosclerosis, vasculitis, heart failure,stroke, myocardial infarction, myocardial ischemia-reperfusion injury,sexual dysfunction, obesity, hypertension, diabetes mellitus anddiabetic complication, hyperlipidemia, preeclampsia, kidney disease,liver disease, kidney injury, liver injury, snake bite, allograftrejection, organ transplantation, graft versus host disease, dementia,Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis,pain, central nervous system disease, uveitis, Behcet's disease,diabetic macular edema, macular degeneration, orbitopathy, glaucoma,hidradenitis suppurativa, multicentric reticulohistiocytosis, pityriasisrubra pilaris, eosinophilic fascilitis, panniculitis, necrobiosislipoidica diabeticorum, cicatricial pemphigoid, pyoderma gangrenosum,Sweet's syndrome, subcorneal pustular dermatosis, scleroderma,neutrophilic dermatitis, toxic epidermal necrolysis, pustulardermatitis, dermatomyositis, polymyositis, bullous dermatosis, erythemanodosum, alopecia, depressive disorder, bipolar disorder, anxietydisorder, tuberculosis, viral infection, bacterial infection, fungalinfection, protozoan infection, cerebral malaria, sepsis, septic shock,prostate cancer, skin cancer, colorectal cancer, kidney cancer,pancreatic cancer, ovarian cancer, breast cancer, bladder cancer,prostate cancer, lymphoma, glioma, osteosarcoma, leukemia, multiplemyeloma, and cachexia, but is not limited thereto.

As used herein, the term “pharmaceutically effective amount” refers toan amount sufficient for a drug to be administered to an animal or ahuman to exhibit the desired physiological or pharmacological activity.However, the pharmaceutically effective amount may be appropriatelychanged depending on the age, body weight, health condition, and sex ofthe subject to be administered, administration routes, and the durationof treatment and the like.

In addition, as used herein, the term “pharmaceutically acceptable”refers to being physiologically acceptable and does not customarilycause an allergic reaction such as gastrointestinal disorder, dizzinessand the like or a reaction similar thereto when administered to a human.Examples of the carrier, excipient, and diluent include lactose,dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol,starch, acacia gum, alginate, gelatin, calcium phosphate, calciumsilicate, cellulose, methyl cellulose, polyvinylpyrrolidone, water,methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesiumstearate, and mineral oil. In addition, it may further comprise afiller, an anticoagulant, a lubricant, a wetting agent, a flavoringagent, an emulsifying agent, and a preservative and the like.

In addition, the present invention provides a reagent composition forinhibiting a TNF activity in vitro, containing a 4-benzopyranonederivative represented by Formula 1, a pharmaceutically acceptable salt,solvate, racemate, or stereoisomer thereof as an active ingredient:

in which,

R¹ is heteroaryl or substituted phenyl;

R² is hydrogen or lower alkyl of (C1-C4); and

R³, R⁴, R⁵, and R⁶ are each independently hydrogen, halogen, or(C1-C4)alkoxy.

Furthermore, the present invention provides a method for inhibiting aTNF activity, comprising a step of treating an animal excluding a humanwith a 4-benzopyranone derivative represented by Formula 1 above, apharmaceutically acceptable salt, solvate, racemate, or stereoisomerthereof.

Specifically, in Formula 1 above, R¹ may be furanyl, thiophenyl, orsubstituted phenyl represented by

and R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ may be each independentlyhydrogen, chloro, bromo, fluoro, or methoxy.

More specifically, the 4-benzopyranone derivative may be selected fromthe group consisting of2-(2-chlorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl) carboxy-8-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy chloro-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl) carboxy-7-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2,5-difluorophenyl) carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl) carboxy-6-chloro-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone; and2-(4-chlorophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone.

EMBODIMENT FOR WORKING THE INVENTION

Hereinafter, the present invention will be described in more detailthrough the working examples in order to promote understanding of thepresent invention. However, the following examples are provided for abetter understanding of the present invention to one of ordinary skillin the art, and are only illustrations of the present invention, but thescope of the present invention is not limited to the following examples.

<Preparation Example 1> General Reaction Process of 4-BenzopyranoneCompound Derivative

First, a 2-hydroxybenzophosphorane derivative compound (2) was dissolvedin carbon dichloride, and pyridine (2 eq) was added dropwise thereto.Then, a heteroaromatic or benzoyl chloride derivative (1.5 eq) was addeddropwise, and 4-dimethylaminopyridine (0.05 eq) was added dropwisethereto. Then, the reaction was stirred under a nitrogen atmosphere for1 hour to 5 hours. When the reaction was completed, acetic acid (1 mL)was added to the reaction solution, and Girard's reagent T (1 eq) wasadded dropwise, and then the mixture was stirred under a nitrogenatmosphere for 1 hour. The mixture was extracted with 1N HCl andsaturated NaHCO₃ solution. The organic layer was dried over anhydrousMgSO₄, and then filtered and distilled under reduced pressure to obtainthe compound (3). The compound (3) was dissolved in a solvent toluene.After refluxing with heating under a nitrogen atmosphere for 5 to 18hours, when the reaction was completed, the solvent was removed bydistillation under reduced pressure, and then the reaction was purifiedby chromatography. The thus obtained 4-benzopyranone compound (4) wasdissolved in carbon dichloride, and TFA was added dropwise, and themixture was stirred under a nitrogen atmosphere for 1 hour to 5 hours.When the reaction was completed, the solvent was removed by distillationunder reduced pressure. The crystals obtained by crystallization withether were filtered and then dried in a vacuum oven to obtain the finaldesired compounds (5).

<Example 1> Synthesis of2-(3-fluorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methoxybenzophosphorane compound (150 mg, 0.28 mmol),pyridine (0.05 mL, 0.57 mmol, 2 eq), and 3-fluorobenzoyl chloride (0.05mL, 0.43 mmol, 1.5 eq) were stirred in carbon dichloride (5 mL) for 4hours and 30 minutes, and stirred in toluene (10 mL) for 2 hours and 30minutes, and then stirred in TFA (2 mL) and carbon dichloride (2 mL) for1 hour and 30 minutes to obtain 60 mg (67.0%) of the final desiredcompound, 2-(3-fluorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone.

m.p. 214-215° C. ¹H NMR (300 MHz, CDCl₃): δ 3.96 (s, 3H), 6.97 (d, J=2.3Hz, 1H), 7.15 (dd, J=9.1, 2.3 Hz, 1H), 7.26-7.50 (m, 4H), 8.24 (d, J=9.1Hz, 1H); ¹³C NMR (75 MHz, CDCl₃): δ 56.22, 100.21, 110.10, 115.48,116.14, 116.91, 118.53, 124.96, 127.77, 129.86, 134.56, 157.54, 160.45,163.56, 166.12, 172.04, 179.62.

<Example 2> Synthesis of2-(2-furanyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methoxybenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 2-furoyl chloride(0.029 mL, 0.30 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 2 hours, and stirred in toluene(6 mL) for 50 hours, and then stirred in TFA (3 mL) and methylenechloride (3 mL) for 1 hour to obtain 25.2 mg (47.6%) of the finaldesired compound,2-(2-furanyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone.

m.p. 195-197° C. ¹H NMR (300 MHz, CDCl₃): δ 2.53 (s, 3H), 6.71 (dd,J=3.7, 1.7 Hz, 1H), 7.55 (d, J=8.6 Hz, 1H), 7.66 (dd, J=8.6, 2.0 Hz,1H), 7.79 (d, J=1.7 Hz, 1H), 7.99 (d, J=3.7 Hz, 1H), 8.09 (s, 1H), 14.55(s, 1H).

<Example 3> Synthesis of2-(2-furanyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methoxybenzophosphorane compound (200 mg, 0.38 mmol),pyridine (0.06 mL, 0.76 mmol, 2 eq), and 2-furoyl chloride (0.05 mL,0.57 mmol, 1.5 eq) were stirred in carbon dichloride (4 mL) for 1 hour,and stirred in toluene (15 mL) for 10 hours, and then stirred in TFA (2mL) and carbon dichloride (2 mL) for 4 hours to obtain 50 mg (46.0%) ofthe final desired compound,2-(2-furanyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone.

m.p. 217-219° C. ¹H NMR (300 MHz, CDCl₃): δ 3.99 (s, 3H), 6.71 (dd,J=3.7, 1.6 Hz, 1H), 7.03 (d, J=2.3 Hz, 1H), 7.11 (dd, J=9.0, 2.3 Hz,1H), 7.78 (d, J=1.6 Hz, 1H), 8.01 (d, J=3.7 Hz, 1H), 8.21 (d, J=9.0 Hz,1H), 15.00 (s, 1H).

<Example 4> Synthesis of2-(2-furanyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methoxybenzophosphorane compound (200 mg, 0.38 mmol),pyridine (0.06 mL, 0.76 mmol, 2 eq), and 2-furoyl chloride (0.05 mL,0.57 mmol, 1.5 eq) were stirred in methylene chloride (4 mL) for 1 hour,and stirred in toluene (15 mL) for 10 hours, and then stirred in TFA (2mL) and methylene chloride (2 mL) for 4 hours to obtain 70 mg (64.0%) ofthe final desired compound,2-(2-furanyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone.

m.p. 205-207° C. ¹H NMR (300 MHz, CDCl₃): δ 3.97 (s, 3H), 6.71 (dd,J=3.7, 1.7 Hz, 1H), 7.43 (dd, J=9.2, 3.1 Hz, 1H), 7.60 (d, J=9.2 Hz,1H), 7.64 (d, J=3.1 Hz, 1H), 7.79 (d, J=1.7 Hz, 1H), 7.99 (d, J=3.7 Hz,1H), 14.60 (s, 1H).

<Example 5> Synthesis of2-(3-methylphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methoxybenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), m-toluoyl chloride(0.031 mL, 0.24 mmol, 1.2 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 24 hours, and stirred intoluene (6 mL) for 14 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour and 30 minutes to obtain 23.1 mg(40.1%) of the final desired compound,2-(3-methylphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone.

m.p. 192-195° C. ¹H NMR (300 MHz, CDCl₃): δ 2.46 (s, 3H), 2.55 (s, 3H),7.41-7.47 (m, 4H), 7.52 (d, J=8.6 Hz, 1H), 7.67 (dd, J=8.6, 2.1 Hz, 1H),8.14 (d, J=0.9 Hz, 1H), 14.30 (s, 1H).

<Example 6> Synthesis of2-(3-methylphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methoxybenzophosphorane compound (200 mg, 0.38 mmol),pyridine (0.06 mL, 0.76 mmol, 2 eq), and m-toluoyl chloride (0.05 mL,0.57 mmol, 1.5 eq) were stirred in methylene chloride (4 mL) for 19hours, and stirred in toluene (15 mL) for 17 hours, and then stirred inTFA (5 mL) and methylene chloride (5 mL) for 2 hours to obtain 76.3 mg(65.0%) of the final desired compound,2-(3-methylphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone.

m.p. 183-184° C. ¹H NMR (300 MHz, CDCl₃): δ 2.46 (s, 3H), 3.96 (s, 3H),6.98 (d, J=2.3 Hz, 1H), 7.14 (dd, J=9.0, 2.3 Hz, 1H), 7.40-7.46 (m, 4H),8.25 (d, J=9.0 Hz, 1H), 14.45 (s, 1H).

<Example 7> Synthesis of2-(3-methylphenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methoxybenzophosphorane compound (200 mg, 0.38 mmol),pyridine (0.06 mL, 0.76 mmol, 2 eq), and m-toluoyl chloride (0.05 mL,0.57 mmol, 1.5 eq) were stirred in methylene chloride (4 mL) for 2hours, and stirred in toluene (15 mL) for 71 hours, and then stirred inTFA (5 mL) and methylene chloride (5 mL) for 2 hours to obtain 49.5 mg(42.0%) of the final desired compound,2-(3-methylphenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone.

m.p. 198-199° C. ¹H NMR (300 MHz, CDCl₃): δ 2.46 (s, 3H), 3.98 (s, 3H),7.27-7.46 (m, 5H), 7.56 (d, J=9.2 Hz, 1H), 7.67 (d, J=3.0 Hz, 1H), 14.35(s, 1H).

<Example 8> Synthesis of2-(3,4-dimethoxyphenyl)-3-carboxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxybenzophosphorane compound (200 mg, 0.4 mmol), pyridine (0.05mL, 0.6 mmol, 1.5 eq), and 3,4-dimethoxybenzoyl chloride (96 mg, 0.48mmol, 1.2 eq) were stirred in methylene chloride (4 mL) for 150 hours,and stirred in toluene (15 mL) for 72 hours, and then stirred in TFA (5mL) and methylene chloride (5 mL) for 2 hours to obtain 34.9 mg (63.0%)of the final desired compound,2-(3,4-dimethoxyphenyl)-3-carboxy-(4H)-4-benzopyranone.

m.p. 237-238° C. ¹H NMR (300 MHz, CDCl₃): δ 3.94 (s, 3H), 3.99 (s, 3H),7.00 (d, J=8.4 Hz, 1H), 7.23 (d, J=2.1 Hz, 1H), 7.39 (dd, J=8.4, 2.1 Hz,1H), 7.56-7.64 (m, 2H), 7.64-7.90 (m, 1H), 8.36 (dd, J=8.0, 1.6 Hz, 1H),14.50 (s, 1H).

<Example 9> Synthesis of2-(3,4-dimethoxyphenyl)-3-carboxy-7-methoxy-(4H)

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methoxybenzophosphorane compound (200 mg, 0.38 mmol),pyridine (0.05 mL, 0.57 mmol, 1.5 eq), and 3,4-dimethoxybenzoyl chloride(92 mg, 0.46 mmol, 1.2 eq) were stirred in methylene chloride (4 mL) for12 hours, and stirred in benzene (10 mL) for 45 hours, and then stirredin TFA (5 mL) and methylene chloride (5 mL) for 2 hours to obtain 59.7mg (89.0%) of the final desired compound,2-(3,4-dimethoxyphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone.

m.p. 208-210° C. ¹H NMR (300 MHz, CDCl₃): δ 3.94 (s, 3H), 3.96 (s, 3H),3.97 (s, 3H), 6.97-6.99 (m, 2H), 7.12 (dd, J=8.9, 2.3 Hz, 1H), 7.20 (d,J=1.9 Hz, 1H), 7.34 (dd, J=8.4, 1.9 Hz, 1H), 8.22 (d, J=8.9 Hz, 1H).

<Example 10> Synthesis of2-(2-chlorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methoxybenzophosphorane compound (200 mg, 0.38 mmol),pyridine (0.05 mL, 0.57 mmol, 1.5 eq), and 2-chlorobenzoyl chloride(0.06 mL, 0.46 mmol, 1.2 eq) were stirred in methylene chloride (4 mL)for 1 hour, and stirred in toluene (15 mL) for 8 hours, and then stirredin TFA (5 mL) and methylene chloride (5 mL) for 2 hours to obtain 70 mg(88.0%) of the desired compound,2-(2-chlorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone.

m.p. 130-132° C. ¹H NMR (300 MHz, CDCl₃): δ 3.96 (s, 3H), 6.98 (d, J=2.4Hz, 1H), 7.17 (dd, J=9.0, 2.4 Hz, 1H), 7.43-7.44 (m, 2H), 7.48-7.56 (m,2H), 8.28 (d, J=9.0 Hz, 1H), 14.40 (s, 1H).

<Example 11> Synthesis of2-(2-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxybenzophosphorane compound (200 mg, 0.40 mmol), pyridine (0.05mL, 0.60 mmol, 1.5 eq), and 2-chlorobenzoyl chloride (0.06 mL, 0.48mmol, 1.2 eq) were stirred in methylene chloride (4 mL) for 1 hour, andstirred in toluene (15 mL) for 120 hours, and then stirred in TFA (5 mL)and methylene chloride (5 mL) for 6 hours and 30 minutes to obtain 34 mg(44.0%) of the final desired compound,2-(2-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone.

m.p. 133-135° C. ¹H NMR (300 MHz, CDCl₃): δ 7.44-7.46 (m, 2H), 7.49-7.55(m, 2H), 7.61-7.66 (m, 2H), 7.87-7.93 (m, 1H), 8.40 (dd, J=8.3, 1.7 Hz,1H).

<Example 12> Synthesis of2-(2-thiophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methoxybenzophosphorane compound (200 mg, 0.38 mmol),pyridine (0.05 mL, 0.57 mmol, 1.5 eq), and 2-thiophenecarbonyl chloride(0.05 mL, 0.46 mmol, 1.2 eq) were stirred in methylene chloride (4 mL)for 2 hours and 30 minutes, and stirred in toluene (15 mL) for 36 hours,and then stirred in TFA (5 mL) and methylene chloride (5 mL) for 2 hoursto obtain 52.4 mg (56.0%) of the final desired compound,2-(2-thiophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone.

m.p. 196-198° C. ¹H NMR (300 MHz, CDCl₃): δ 4.07 (s, 3H), 7.25 (dd,J=5.0, 4.0 Hz, 1H), 7.31 (dd, J=8.1, 1.3 Hz, 1H), 7.46 (dd, J=8.1, 8.1Hz, 1H), 7.81-7.85 (m, 2H), 8.38 (dd, J=4.0, 1.2 Hz, 1H), 14.80 (s, 1H).

<Example 13> Synthesis of2-(2-thiophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methoxybenzophosphorane compound (200 mg, 0.38 mmol),pyridine (0.05 mL, 0.57 mmol, 1.5 eq), and 2-thiophenecarbonyl chloride(0.05 mL, 0.46 mmol, 1.2 eq) were stirred in methylene chloride (4 mL)for 2 hours and 30 minutes, and stirred in toluene (15 mL) for 28 hours,and then stirred in TFA (5 mL) and methylene chloride (5 mL) for 2 hoursto obtain 41 mg (77.0%) of the final desired compound,2-(2-thiophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone.

m.p. 205-207° C. ¹H NMR (300 MHz, CDCl₃): δ 3.97 (s, 3H), 6.95 (d, J=2.3Hz, 1H), 7.09 (dd, J=9.0, 2.3 Hz, 1H), 7.22 (dd, J=5.0, 4.0 Hz, 1H),7.79 (dd, J=5.0, 1.1 Hz, 1H), 8.19 (d, J=9.0 Hz, 1H), 8.25 (dd, J=4.0,1.1 Hz, 1H), 14.98 (s, 1H).

<Example 14> Synthesis of2-(2-thiophenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methoxybenzophosphorane compound (200 mg, 0.38 mmol),pyridine (0.05 mL, 0.57 mmol, 1.5 eq), and 2-thiophenecarbonyl chloride(0.05 mL, 0.46 mmol, 1.2 eq) were stirred in methylene chloride (4 mL)for 2 hours and 30 minutes, and stirred in toluene (15 mL) for 42 hours,and then stirred in TFA (5 mL) and methylene chloride (5 mL) for 1 hourand 30 minutes to obtain 50 mg (68.0%) of the final desired compound,2-(2-thiophenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone.

m.p. 172-174° C. ¹H NMR (300 MHz, CDCl₃): δ 3.96 (s, 3H), 7.24 (dd,J=5.0, 4.0 Hz, 1H), 7.43 (dd, J=9.2, 3.0 Hz, 1H), 7.55 (d, J=9.2 Hz,1H), 7.63 (d, J=3.0 Hz, 1H), 7.81 (dd, J=5.0, 1.2 Hz, 1H), 8.25 (dd,J=4.0, 1.2 Hz, 1H), 14.78 (s, 1H).

<Example 15> Synthesis of2-(2-thiophenyl)-3-carboxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxybenzophosphorane compound (200 mg, 0.40 mmol), pyridine (0.05mL, 0.60 mmol, 1.5 eq), and 2-thiophenecarbonyl chloride (0.05 mL, 0.48mmol, 1.2 eq) were stirred in methylene chloride (4 mL) for 3 hours, andstirred in toluene (15 mL) for 23 hours and 30 minutes, and then stirredin TFA (5 mL) and methylene chloride (5 mL) for 2 hours to obtain 52.3mg (78.0%) of the final desired compound,2-(2-thiophenyl)-3-carboxy-(4H)-4-benzopyranone.

m.p. 175-176° C. ¹H NMR (300 MHz, CDCl₃): δ 7.25 (dd, J=5.0, 4.1 Hz,1H), 7.55-7.64 (m, 2H), 7.83-7.89 (m, 2H), 8.29-8.34 (m, 2H), 14.70 (s,1H).

<Example 16> Synthesis of2-(3-fluorophenyl)-3-carboxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxybenzophosphorane compound (300 mg, 0.6 mmol),diisopropylethylamine (0.314 mL, 1.8 mmol, 3 eq), 3-fluorobenzoylchloride (0.118 mL, 1.2 mmol, 2 eq), and 4-(dimethylamino)pyridine (10mg) were stirred in methylene chloride (5 mL) for 1 hour, and stirred intoluene (15 mL) for 4 hours and 30 minutes, and then stirred in TFA (7mL) and methylene chloride (7 mL) for 1 hour and 30 minutes to obtain154 mg (90.0%) of the final desired compound,2-(3-fluorophenyl)-3-carboxy-(4H)-4-benzopyranone.

m.p. 193-196° C. ¹H NMR (300 MHz, CDCl₃): δ 7.29-7.55 (m, 4H), 7.60-7.65(m, 2H), 7.87-7.93 (m, 1H), 8.37-8.40 (m, 1H), 14.19 (s, 1H).

<Example 17> Synthesis of2-(3,5-difluorophenyl)-3-carboxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxybenzophosphorane compound (300 mg, 0.6 mmol),diisopropylethylamine (0.314 mL, 1.8 mmol, 3 eq), 3,5-difluorobenzoylchloride (0.15 mL, 1.2 mmol, 2 eq), and 4-(dimethylamino)pyridine (10mg) were stirred in methylene chloride (5 mL) for 1 hour, and stirred intoluene (15 mL) for 2 hours and 30 minutes, and then stirred in TFA (7mL) and methylene chloride (7 mL) for 1 hour to obtain 22 mg (12.0%) ofthe final desired compound,2-(3,5-difluorophenyl)-3-carboxy-(4H)-4-benzopyranone.

m.p. 208-209° C. ¹H NMR (300 MHz, CDCl₃): δ 7.02-7.08 (m, 1H), 7.16-7.19(m, 2H), 7.62-66 (m, 2H), 7.89-7.94 (m, 1H), 8.38 (d, J=8.3 Hz, 1H).

<Example 18> Synthesis of 2-(2-furanyl)-3-carboxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxybenzophosphorane compound (300 mg, 0.6 mmol),diisopropylethylamine (0.314 mL, 1.8 mmol, 3 eq), 2-furoyl chloride(0.118 mL, 1.2 mmol, 2 eq), and 4-(dimethylamino)pyridine (10 mg) werestirred in methylene chloride (5 mL) for 1 hour, and stirred in toluene(15 mL) for 8 hours, and then stirred in TFA (7 mL) and methylenechloride (7 mL) for 1 hour to obtain 96 mg (62.0%) of the final desiredcompound, 2-(2-furanyl)-3-carboxy-(4H)-4-benzopyranone.

m.p. 202-205° C. ¹H NMR (300 MHz, CDCl₃): δ 6.72 (dd, J=3.8, 1.7 Hz,1H), 7.54-7.60 (m, 1H), 7.65 (d, J=8.4 Hz, 1H), 7.81-7.89 (m, 2H),8.01-8.03 (m, 1H), 8.32 (dd, J=8.0, 1.5 Hz, 1H), 14.45 (s, 1H).

<Example 19> Synthesis of2-(3-methylphenyl)-3-carboxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxybenzophosphorane compound (300 mg, 0.6 mmol),diisopropylethylamine (0.314 mL, 1.8 mmol, 3 eq), m-toluoyl chloride(0.158 mL, 1.2 mmol, 2 eq), and 4-(dimethylamino)pyridine (10 mg) werestirred in methylene chloride (5 mL) for 1 hour, and stirred in toluene(15 mL) for 72 hours, and then stirred in TFA (7 mL) and methylenechloride (7 mL) for 1 hour to obtain 73 mg (43.0%) of the final desiredcompound, 2-(3-methylphenyl)-3-carboxy-(4H)-4-benzopyranone.

m.p. 162-164° C. ¹H NMR (300 MHz, CDCl₃): δ 2.48 (s, 3H), 7.43-7.50 (m,4H), 7.59-7.65 (m, 2H), 7.86-7.92 (m, 1H), 8.38 (dd, J=8.0, 1.5 Hz, 1H),14.20 (s, 1H).

<Example 20> Synthesis of2-(2-furanyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 2-furoyl chloride(0.029 mL, 0.30 mmol, 1.5 eq), 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 2 hours, and stirred in toluene(6 mL) for 50 hours, and then stirred in TFA (3 mL) and methylenechloride (3 mL) for 1 hour to obtain 31.4 mg (59.3%) of the finaldesired compound, 2-(2-furanyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone.

m.p. 227-228° C. ¹H NMR (300 MHz, CDCl₃): δ 2.58 (s, 3H), 6.69 (dd,J=3.6, 1.7 Hz, 1H), 7.32-7.36 (m, 2H), 7.59 (d, J=7.3 Hz, 1H), 7.76-7.77(m, 1H), 7.98 (d, J=7.3 Hz, 1H).

<Example 21> Synthesis of2-(2-furanyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 2-furoyl chloride(0.029 mL, 0.30 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 2 hours, and stirred in toluene(6 mL) for 50 hours, and then stirred in TFA (3 mL) and methylenechloride (3 mL) for 1 hour to obtain 19.1 mg (36.1%) of the finaldesired compound, 2-(2-furanyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone.

m.p. 217-218° C. ¹H NMR (300 MHz, CDCl₃): δ 2.57 (s, 3H), 6.71 (dd,J=3.7, 1.7 Hz, 1H), 7.37 (dd, J=8.2, 0.8 Hz, 1H), 7.46 (s, 1H), 7.79 (d,J=1.7 Hz, 1H), 8.00 (d, J=3.7 Hz, 1H), 8.18 (d, J=8.2 Hz, 1H), 14.55 (s,1H).

<Example 22> Synthesis of2-(2-furanyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 2-furoyl chloride(0.029 mL, 0.30 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 2 hours, and stirred in toluene(6 mL) for 50 hours, and then stirred in TFA (3 mL) and methylenechloride (3 mL) for 1 hour to obtain 25.2 mg (47.6%) of the finaldesired compound, 2-(2-furanyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone.

m.p. 195-197° C. ¹H NMR (300 MHz, CDCl₃): δ 2.53 (s, 3H), 6.71 (dd,J=3.7, 1.7 Hz, 1H), 7.55 (d, J=8.6 Hz, 1H), 7.66 (dd, J=8.6, 2.0 Hz,1H), 7.79 (d, J=1.7 Hz, 1H), 7.99 (d, J=3.7 Hz, 1H), 8.09 (s, 1H), 14.55(s, 1H).

<Example 23> Synthesis of2-(2-furanyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-chlorobenzophosphorane compound (100 mg, 0.19 mmol),diisopropylethylamine (0.066 mL, 0.38 mmol, 2 eq), 2-furoyl chloride(0.028 mL, 0.29 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 2 hours, and stirred in toluene(6 mL) for 50 hours, and then stirred in TFA (3 mL) and methylenechloride (3 mL) for 1 hour to obtain 20.20 mg (36.9%) of the finaldesired compound, 2-(2-furanyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone.

m.p. 196-198° C. ¹H NMR (300 MHz, CDCl₃): δ 6.73 (dd, J=3.8, 1.7 Hz,1H), 7.62 (d, J=9.0 Hz, 1H), 7.77-7.81 (m, 2H), 8.04 (dd, J=3.8, 0.5 Hz,1H), 8.27 (d, J=2.5 Hz, 1H), 14.00 (s, 1H).

<Example 24> Synthesis of2-(2-furanyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-bromobenzophosphorane compound (100 mg, 0.17 mmol),diisopropylethylamine (0.061 mL, 0.34 mmol, 2 eq), 2-furoyl chloride(0.026 mL, 0.26 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 2 hours, and stirred in toluene(6 mL) for 50 hours, and then stirred in TFA (3 mL) and methylenechloride (3 mL) for 1 hour to obtain 41.9 mg (71.9%) of the finaldesired compound, 2-(2-furanyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone.

m.p. 209-211° C. ¹H NMR (300 MHz, CDCl₃): δ 6.73 (dd, J=3.8, 1.6 Hz,1H), 7.55 (d, J=8.9 Hz, 1H), 7.80-7.81 (m, 1H), 7.93 (dd, J=8.9, 2.4 Hz,1H), 8.03 (d, J=3.8 Hz, 1H), 8.43 (d, J=2.4 Hz, 1H), 14.25 (s, 1H).

<Example 25> Synthesis of2-(3-methylphenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), m-toluoyl chloride(0.031 mL, 0.24 mmol, 1.2 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 24 hours, and stirred intoluene (6 mL) for 144 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour and 30 minutes to obtain 14.0 mg(24.3%) of the final desired compound,2-(3-methylphenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone.

m.p. 209-211° C. ¹H NMR (300 MHz, CDCl₃): δ 2.47 (s, 3H), 2.55 (s, 3H),7.42-7.51 (m, 5H), 7.70 (d, J=7.1 Hz, 1H), 8.19 (d, J=7.1 Hz, 1H), 14.25(s, 1H).

<Example 26> Synthesis of2-(3-methylphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), m-toluoyl chloride(0.031 mL, 0.24 mmol, 1.2 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 24 hours, and stirred intoluene (6 mL) for 144 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour and 30 minutes to obtain 23.2 mg(40.4%) of the final desired compound,2-(3-methylphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone.

m.p. 152-155° C. ¹H NMR (300 MHz, CDCl₃): δ 2.46 (s, 3H), 2.57 (s, 3H),7.39-7.47 (m, 6H), 8.24 (d, J=8.5 Hz, 1H), 14.30 (s, 1H).

<Example 27> Synthesis of2-(3-methylphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), m-toluoyl chloride(0.031 mL, 0.24 mmol, 1.2 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 24 hours, and stirred intoluene (6 mL) for 144 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour and 30 minutes to obtain 23.1 mg(40.1%) of the final desired compound,2-(3-methylphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone.

m.p. 192-195° C. ¹H NMR (300 MHz, CDCl₃): δ 2.46 (s, 3H), 2.55 (s, 3H),7.41-7.47 (m, 4H), 7.52 (d, J=8.6 Hz, 1H), 7.67 (dd, J=8.6, 2.1 Hz, 1H),8.14 (d, J=0.9 Hz, 1H), 14.30 (s, 1H).

<Example 28> Synthesis of2-(3-methylphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-chlorobenzophosphorane compound (100 mg, 0.19 mmol),diisopropylethylamine (0.066 mL, 0.38 mmol, 2 eq), m-toluoyl chloride(0.030 mL, 0.23 mmol, 1.2 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 24 hours, and stirred intoluene (6 mL) for 144 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour and 30 minutes to obtain 28.4 mg(48.0%) of the final desired compound,2-(3-methylphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone.

m.p. 179-181° C. ¹H NMR (300 MHz, CDCl₃): δ 2.46 (s, 3H), 7.43-7.46 (m,4H), 7.56 (dd, J=8.6, 1.5 Hz, 1H), 7.66 (s, 1H), 8.30 (d, J=8.6 Hz, 1H),13.95 (s, 1H).

<Example 29> Synthesis of2-(3-methylphenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-chlorobenzophosphorane compound (100 mg, 0.19 mmol),diisopropylethylamine (0.066 mL, 0.38 mmol, 2 eq), m-toluoyl chloride(0.030 mL, 0.23 mmol, 1.2 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 24 hours, and stirred intoluene (6 mL) for 144 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour and 30 minutes to obtain 13.4 mg(22.6%) of the final desired compound,2-(3-methylphenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone.

m.p. 218-220° C. ¹H NMR (300 MHz, CDCl₃): δ 2.46 (s, 3H), 7.44-7.47 (m,4H), 7.52 (d, J=8.9 Hz, 1H), 7.95 (dd, J=8.9, 2.4 Hz, 1H), 8.49 (d,J=2.4 Hz, 1H), 13.80 (s, 1H).

<Example 30> Synthesis of2-(3-methylphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-bromobenzophosphorane compound (100 mg, 0.17 mmol),diisopropylethylamine (0.061 mL, 0.34 mmol, 2 eq), m-toluoyl chloride(0.027 mL, 0.20 mmol, 1.2 eq), and 4-(dimethylamino)pyridine (5 mg) werestirred in methylene chloride (3 mL) for 24 hours, and stirred intoluene (6 mL) for 144 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour and 30 minutes to obtain 21.6 mg(34.6%) of the final desired compound,2-(3-methylphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone.

m.p. 208-210° C. ¹H NMR (300 MHz, CDCl₃): δ 2.47 (s, 3H), 7.42-7.47 (m,4H), 7.59 (d, J=8.9 Hz, 1H), 7.80 (dd, J=8.9, 2.5 Hz, 1H), 8.32 (d,J=2.5 Hz, 1H), 13.90 (s, 1H).

<Example 31> Synthesis of2-(2-thiophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 2-thiophenecarbonylchloride (0.031 mL, 0.30 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 1 hour, and stirred intoluene (6 mL) for 21 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour to obtain 26.5 mg (47.3%) of thefinal desired compound,2-(2-thiophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone.

m.p. 205-206° C. ¹H NMR (300 MHz, CDCl₃): δ 2.63 (s, 3H), 7.25-7.26 (m,4H), 7.45 (dd, J=7.7, 7.7 Hz, 1H), 7.68 (d, J=7.7 Hz, 1H), 7.83 (dd,J=5.0, 1.0 Hz, 1H), 8.14 (d, J=7.7 Hz, 1H), 8.38 (dd, J=4.0, 1.0 Hz,1H).

<Example 32> Synthesis of2-(2-thiophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 2-thiophenecarbonylchloride (0.031 mL, 0.30 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 1 hour, and stirred intoluene (6 mL) for 21 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour to obtain 30.1 mg (53.8%) of thefinal desired compound,2-(2-thiophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone.

m.p. 202-203° C. ¹H NMR (300 MHz, CDCl₃): δ 2.58 (s, 3H), 7.24 (dd,J=5.0, 4.0 Hz, 1H), 7.37 (d, J=8.2 Hz, 1H), 7.42 (s, 1H), 7.82 (dd,J=5.0, 1.1 Hz, 1H), 8.18 (d, J=8.2 Hz, 1H), 8.28 (dd, J=4.0, 1.1 Hz,1H), 14.80 (s, 1H).

<Example 33> Synthesis of2-(2-thiophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 2-thiophenecarbonylchloride (0.031 mL, 0.30 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 1 hour, and stirred intoluene (6 mL) for 30 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour to obtain 50 mg (89%) of the finaldesired compound,2-(2-thiophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone.

m.p. 153-155° C. ¹H NMR (300 MHz, CDCl₃): δ 2.54 (s, 3H), 7.24 (dd,J=5.0, 4.0 Hz, 1H), 7.52 (d, J=8.6 Hz, 1H), 7.66 (dd, J=8.6, 2.0 Hz,1H), 7.82 (dd, J=5.0, 1.1 Hz, 1H), 8.09 (s, 1H), 8.28 (dd, J=4.0, 1.1Hz, 1H), 14.75 (s, 1H).

<Example 34> Synthesis of2-(2-thiophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-chlorobenzophosphorane compound (100 mg, 0.19 mmol),diisopropylethylamine (0.066 mL, 0.38 mmol, 2 eq), 2-thiophenecarbonylchloride (0.030 mL, 0.29 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 1 hour, and stirred intoluene (6 mL) for 24 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour to obtain 37.0 mg (63.8%) of thefinal desired compound,2-(2-thiophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone.

m.p. 232-233° C. ¹H NMR (300 MHz, CDCl₃): δ 7.20 (dd, J=5.0, 4.0 Hz,1H), 7.42 (dd, J=8.5, 1.8 Hz, 1H), 7.60 (d, J=1.8 Hz, 1H), 7.73 (dd,J=5.0, 1.0 Hz, 1H), 7.93 (dd, J=4.0, 1.0 Hz, 1H), 8.15 (d, J=8.5 Hz,1H).

<Example 35> Synthesis of2-(2-thiophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-chlorobenzophosphorane compound (100 mg, 0.19 mmol),diisopropylethylamine (0.066 mL, 0.38 mmol, 2 eq), 2-thiophenecarbonylchloride (0.030 mL, 0.29 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 1 hour, and stirred intoluene (6 mL) for 72 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour to obtain 36.7 mg (63.3%) of thefinal desired compound,2-(2-thiophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone.

m.p. 202-204° C. ¹H NMR (300 MHz, CDCl₃): δ 7.24 (dd, J=5.0, 4.0 Hz,1H), 7.57 (d, J=8.9 Hz, 1H), 7.77 (dd, J=8.9, 2.5 Hz, 1H), 7.84 (dd,J=5.0, 1.1 Hz, 1H), 8.25 (d, J=2.5 Hz, 1H), 8.28 (dd, J=4.0, 1.1 Hz,1H), 14.45 (s, 1H).

<Example 36> Synthesis of2-(2-thiophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-bromobenzophosphorane compound (100 mg, 0.17 mmol),diisopropylethylamine (0.061 mL, 0.34 mmol, 2 eq), 2-thiophenecarbonylchloride (0.028 mL, 0.26 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 1 hour, and stirred intoluene (6 mL) for 72 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour to obtain 46.2 mg (75.7%) of thefinal desired compound,2-(2-thiophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone.

m.p. 210-212° C. ¹H NMR (300 MHz, CDCl₃): δ 7.25 (dd, J=5.0, 4.1 Hz,1H), 7.52 (d, J=8.9 Hz, 1H), 7.86 (dd, J=5.0, 1.1 Hz, 1H), 7.93 (dd,J=8.9, 2.4 Hz, 1H), 8.30 (dd, J=4.1, 1.1 Hz, 1H), 8.44 (d, J=2.4 Hz,1H), 14.30 (s, 1H).

<Example 37> Synthesis of2-(3,4-dimethoxyphenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 3,4-dimethoxybenzoylchloride (59 mg, 0.30 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 24 hours, and stirredin toluene (6 mL) for 96 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour to obtain 20.3 mg (30.3%) of thefinal desired compound,2-(3,4-dimethoxyphenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone.

m.p. 210-212° C. ¹H NMR (300 MHz, CDCl₃): δ 2.58 (s, 3H), 3.96 (s, 3H),4.00 (s, 3H), 7.01 (d, J=8.5 Hz, 1H), 7.27 (d, J=2.1 Hz, 1H), 7.43 (dd,J=8.5, 2.1 Hz, 1H), 7.48 (dd, J=7.7, 7.7 Hz, 1H), 7.69 (d, J=7.7 Hz,1H), 8.18 (d, J=7.7 Hz, 1H), 14.45 (s, 1H).

<Example 38> Synthesis of2-(3,4-dimethoxyphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 3,4-dimethoxybenzoylchloride (59 mg, 0.30 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 24 hours, and stirredin toluene (6 mL) for 96 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour to obtain 28.4 mg (42.4%) of thefinal desired compound,2-(3,4-dimethoxyphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone.

m.p. 200-201° C. ¹H NMR (300 MHz, CDCl₃): δ 2.58 (s, 3H), 3.96 (s, 3H),3.99 (s, 3H), 6.99 (d, J=8.5 Hz, 1H), 7.22 (d, J=2.0 Hz, 1H), 7.36-7.43(m, 3H), 8.22 (d, J=8.1 Hz, 1H), 14.45 (s, 1H).

<Example 39> Synthesis of2-(3,4-dimethoxyphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-chlorobenzophosphorane compound (100 mg, 0.19 mmol),diisopropylethylamine (0.066 mL, 0.38 mmol, 2 eq), 3,4-dimethoxybenzoylchloride (57 mg, 0.29 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 2 hours, and stirredin toluene (6 mL) for 100 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour to obtain 34.8 mg (51.2%) of thefinal desired compound,2-(3,4-dimethoxyphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone.

m.p. 226-227° C. ¹H NMR (300 MHz, CDCl₃): δ 3.95 (s, 3H), 3.99 (s, 3H),6.99 (d, J=8.4 Hz, 1H), 7.21 (d, J=2.1 Hz, 1H), 7.38 (dd, J=8.4, 2.1 Hz,1H), 7.54 (dd, J=8.6, 1.8 Hz, 1H), 7.66 (dd, J=1.8 Hz, 1H), 8.28 (d,J=8.6 Hz, 1H), 14.10 (s, 1H).

<Example 40> Synthesis of2-(3,4-dimethoxyphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-bromobenzophosphorane compound (100 mg, 0.17 mmol),diisopropylethylamine (0.061 mL, 0.34 mmol, 2 eq), 2-thiophenecarbonylchloride (0.028 mL, 0.26 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 1 hour, and stirred intoluene (6 mL) for 92 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour to obtain 34 mg (48%) of the finaldesired compound,2-(3,4-dimethoxyphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone.

m.p. 242-243° C. ¹H NMR (300 MHz, CDCl₃): δ 3.95 (s, 3H), 3.99 (s, 3H),6.99 (d, J=8.4 Hz, 1H), 7.22 (d, J=2.1 Hz, 1H), 7.38 (dd, J=8.4, 2.1 Hz,1H), 7.53 (d, J=8.9 Hz, 1H), 7.94 (dd, J=8.9, 2.4 Hz, 1H), 8.48 (d,J=2.4 Hz, 1H).

<Example 41> Synthesis of2-(3-fluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 3-fluorobenzoylchloride (0.036 mL, 0.30 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 1 hour, and stirred intoluene (6 mL) for 12 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 2 hours to obtain 18.7 mg (32.2%) of thefinal desired compound,2-(3-fluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone.

m.p. 227-229° C. ¹H NMR (300 MHz, CDCl₃): δ 2.54 (s, 3H), 7.28-7.33 (m,1H), 7.39 (d, J=9.3 Hz, 1H), 7.46-7.49 (m. 3H), 7.71 (d, J=7.5 Hz, 1H),8.19 (d, J=8.0 Hz, 1H).

<Example 42> Synthesis of2-(3-fluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 3-fluorobenzoylchloride (0.036 mL, 0.30 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 1 hour, and stirred intoluene (6 mL) for 10 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 2 hours to obtain 16.1 mg (27.8%) of thefinal desired compound,2-(3-fluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone.

m.p. 178-181° C. ¹H NMR (300 MHz, CDCl₃): δ 2.58 (s, 3H), 7.28-7.38 (m,2H), 7.42-7.44 (m, 3H), 7.47-7.54 (m. 1H), 8.23-8.26 (m, 1H), 14.33 (s,1H).

<Example 43> Synthesis of2-(3-fluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methylbenzophosphorane compound (100 mg, 0.20 mmol),diisopropylethylamine (0.068 mL, 0.40 mmol, 2 eq), 3-fluorobenzoylchloride (0.036 mL, 0.30 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 1 hour, and stirred intoluene (6 mL) for 21 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 2 hours to obtain 34.2 mg (59.0%) of thefinal desired compound,2-(3-fluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone.

m.p. 214-215° C. ¹H NMR (300 MHz, CDCl₃): δ 2.56 (s, 3H), 7.30-7.33 (m,1H), 7.35-7.39 (m, 1H), 7.42-7.45 (m, 1H), 7.47-7.54 (m, 2H), 7.69 (dd,J=8.6, 2.0 Hz, 1H), 8.14 (s, 1H), 14.30 (s, 1H).

<Example 44> Synthesis of2-(3-fluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-chlorobenzophosphorane compound (100 mg, 0.19 mmol),diisopropylethylamine (0.066 mL, 0.38 mmol, 2 eq), 3-fluorobenzoylchloride (0.034 mL, 0.29 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 2 hours, and stirredin toluene (6 mL) for 19 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 2 hours to obtain 43 mg (71.6%) of thefinal desired compound,2-(3-fluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone.

m.p. 160-163° C. ¹H NMR (300 MHz, CDCl₃): δ 7.30-7.44 (m, 2H), 7.51 (dd,J=8.0, 5.4 Hz, 1H), 7.58 (dd, J=8.6, 1.8 Hz, 1H), 7.66 (d, J=1.8 Hz,1H), 7.71-7.75 (m, 1H), 8.31 (d, J=8.6 Hz, 1H), 15.35 (s, 1H).

<Example 45> Synthesis of2-(3-fluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-chlorobenzophosphorane compound (100 mg, 0.19 mmol),diisopropylethylamine (0.066 mL, 0.38 mmol, 2 eq), 3-fluorobenzoylchloride (0.034 mL, 0.29 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 2 hours, and stirredin toluene (6 mL) for 24 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 2 hours to obtain 48.0 mg (80.0%) of thefinal desired compound,2-(3-fluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone.

m.p. 216-217° C. ¹H NMR (300 MHz, CDCl₃): δ 7.29-7.39 (m, 2H), 7.42-7.46(m, 1H), 7.49-7.56 (m, 1H), 7.60 (d, J=8.9 Hz, 1H), 7.83 (dd, J=8.9, 2.5Hz, 1H), 8.33 (d, J=2.5 Hz, 1H), 13.80 (s, 1H).

<Example 46> Synthesis of2-(3-fluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-bromobenzophosphorane compound (100 mg, 0.17 mmol),diisopropylethylamine (0.061 mL, 0.34 mmol, 2 eq), 3-fluorobenzoylchloride (0.032 mL, 0.26 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (3 mL) for 1 hour, and stirred intoluene (6 mL) for 24 hours, and then stirred in TFA (3 mL) andmethylene chloride (3 mL) for 1 hour to obtain 44.9 mg (71.3%) of thefinal desired compound,2-(3-fluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone.

m.p. 222-223° C. ¹H NMR (300 MHz, CDCl₃): δ 7.32-7.38 (m, 2H), 7.44 (d,J=7.8 Hz, 1H), 7.48-7.55 (m, 2H), 7.97 (dd, J=9.7, 2.3 Hz, 1H), 8.50 (d,J=2.3 Hz, 1H), 13.85 (s, 1H).

<Example 47> Synthesis of2-(3,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 3,5-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour and 30 minutes,and stirred in toluene (9 mL) for 7 hours, and then stirred in TFA (6mL) and methylene chloride (6 mL) for 2 hours to obtain 92.2 mg (75.0%)of the final desired compound,2-(3,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone.

m.p. 243-244° C. ¹H NMR (300 MHz, CDCl₃): δ 2.53 (s, 3H), 7.01-7.06 (m,1H), 7.26-7.33 (m, 2H), 7.42-7.47 (m. 1H), 7.67 (d, J=7.0 Hz, 1H), 8.13(d, J=7.9 Hz, 1H).

<Example 48> Synthesis of2-(3,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 3,5-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour and 30 minutes,and stirred in toluene (9 mL) for 7 hours, and then stirred in TFA (6mL) and methylene chloride (6 mL) for 2 hours to obtain 77.3 mg (62.9%)of the final desired compound,2-(3,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone.

m.p. 220-222° C. ¹H NMR (300 MHz, CDCl₃): δ 2.65 (s, 3H), 7.07-7.15 (m,1H), 7.18-7.26 (m, 2H), 7.50 (d, J=8.2 Hz, 2H), 8.31 (d, J=8.2 Hz, 1H),14.34 (s, 1H).

<Example 49> Synthesis of2-(3,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methylbenzophosphorane compound (200 mg, 0.40 mml),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 3,5-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour and 30 minutes,and stirred in toluene (9 mL) for 10 hours, and then stirred in TFA (6mL) and methylene chloride (6 mL) for 2 hours to obtain 75.7 mg (61.5%)of the final desired compound,2-(3,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone.

m.p. 238-240° C. ¹H NMR (300 MHz, CDCl₃): δ 2.63 (s, 3H), 7.07-7.15 (m,1H), 7.20-7.34 (m, 2H), 7.59 (d, J=8.6 Hz, 1H), 7.77 (dd, J=8.6, 2.1 Hz,1H), 8.21 (s, 1H), 14.12 (s, 1H).

<Example 50> Synthesis of2-(3,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-chlorobenzophosphorane compound (200 mg, 0.38 mmol),diisopropylethylamine (0.131 mL, 0.75 mmol, 2 eq), 3,5-difluorobenzoylchloride (0.071 mL, 0.57 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 10 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 99.0 mg (78.6%) of thefinal desired compound,2-(3,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone.

m.p. 207-209° C. ¹H NMR (300 MHz, CDCl₃): δ 7.09-7.17 (m, 1H), 7.20-7.12(m, 2H), 7.66 (d, J=8.9 Hz, 1H), 7.91 (dd, J=8.9, 2.5 Hz, 1H), 8.40 (d,J=2.5 Hz, 1H), 14.00 (s, 1H).

<Example 51> Synthesis of2-(3,5-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-bromobenzophosphorane compound (200 mg, 0.35 mmol),diisopropylethylamine (0.121 mL, 0.70 mmol, 2 eq), 3,5-difluorobenzoylchloride (0.066 mL, 0.52 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 10 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 91.3 mg (69.2%) of thefinal desired compound,2-(3,5-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone.

m.p. 216-218° C. ¹H NMR (300 MHz, CDCl₃): δ 7.01-7.08 (m, 1H), 7.14-7.18(m, 2H), 7.51 (d, J=8.9 Hz, 1H), 7.97 (dd, J=8.9, 2.3 Hz, 1H), 8.48 (d,J=2.3 Hz, 1H), 13.60 (s, 1H).

<Example 52> Synthesis of2-(2,3-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 2.3-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), 4-(dimethylamino)pyridine (5 mg)were stirred in methylene chloride (4 mL) for 1 hour and 30 minutes, andstirred in toluene (9 mL) for 15 hours, and then stirred in TFA (6 mL)and methylene chloride (6 mL) for 2 hours to obtain 68.0 mg (55.3%) ofthe final desired compound,2-(2,3-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone.

m.p. 167-169° C. ¹H NMR (300 MHz, CDCl₃): δ 2.51 (s, 3H), 7.26-7.31 (m,2H), 7.36-7.43 (m, 1H), 7.48-7.53 (m, 1H), 7.71 (d, J=7.3 Hz, 1H), 8.20(d, J=7.9 Hz, 1H), 14.21 (s, 1H).

<Example 53> Synthesis of2-(2,3-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 2.3-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour and 30 minutes,and stirred in toluene (9 mL) for 15 hours, and then stirred in TFA (6mL) and methylene chloride (6 mL) for 2 hours to obtain 78.0 mg (63.4%)of the final desired compound,2-(2,3-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone.

m.p. 155-156° C. ¹H NMR (300 MHz, CDCl₃): δ 2.57 (s, 3H), 7.17-7.27 (m,3H), 7.42 (d, J=8.2 Hz, 2H), 8.24 (d, J=8.2 Hz, 1H), 14.20 (s, 1H).

<Example 54> Synthesis of2-(2,3-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 2.3-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour and 30 minutes,and stirred in toluene (9 mL) for 15 hours, and then stirred in TFA (6mL) and methylene chloride (6 mL) for 2 hours to obtain 32.0 mg (26.0%)of the final desired compound,2-(2,3-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone.

m.p. 120-122° C. ¹H NMR (300 MHz, CDCl₃): δ 2.55 (s, 3H), 7.24-7.30 (m,2H), 7.35-7.43 (m, 1H), 7.51 (d, J=8.6 Hz, 1H), 7.69 (dd, J=8.6, 2.0 Hz,1H), 8.14 (s, 1H), 14.15 (s, 1H).

<Example 55> Synthesis of2-(2,3-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-chlorobenzophosphorane compound (200 mg, 0.38 mmol),diisopropylethylamine (0.131 mL, 0.75 mmol, 2 eq), 2.3-difluorobenzoylchloride (0.071 mL, 0.57 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour and 30 minutes,and stirred in toluene (9 mL) for 15 hours, and then stirred in TFA (6mL) and methylene chloride (6 mL) for 2 hours to obtain 26.0 mg (20.6%)of the final desired compound,2-(2,3-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone.

m.p. 220° C. or more. ¹H NMR (300 MHz, CDCl₃): δ 6.95-7.31 (m, 2H),7.38-7.46 (m, 1H), 7.69 (dd, J=8.6, 1.8 Hz, 1H), 7.65 (d, J=1.8 Hz, 1H),8.32 (d, J=8.6 Hz, 1H), 13.85 (s, 1H).

<Example 56> Synthesis of2-(2,3-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-chlorobenzophosphorane compound (200 mg, 0.38 mmol),diisopropylethylamine (0.131 mL, 0.75 mmol, 2 eq), 2.3-difluorobenzoylchloride (0.071 mL, 0.57 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour and 30 minutes,and stirred in toluene (9 mL) for 15 hours, and then stirred in TFA (6mL) and methylene chloride (6 mL) for 2 hours to obtain 34.0 mg (27.0%)of the final desired compound,2-(2,3-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone.

m.p. 120° C. or more. ¹H NMR (300 MHz, CDCl₃): δ 7.24-7.32 (m, 2H),7.38-7.47 (m, 1H), 7.60 (d, J=8.9 Hz, 1H), 7.84 (dd, J=8.9, 2.5 Hz, 1H),8.35 (d, J=2.5 Hz, 1H), 13.81 (s, 1H).

<Example 57> Synthesis of2-(2,3-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-bromobenzophosphorane compound (200 mg, 0.35 mmol),diisopropylethylamine (0.121 mL, 0.70 mmol, 2 eq), 3,5-difluorobenzoylchloride (0.066 mL, 0.52 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour and 30 minutes,and stirred in toluene (9 mL) for 15 hours, and then stirred in TFA (6mL) and methylene chloride (6 mL) for 2 hours to obtain 90.0 mg (68.2%)of the final desired compound,2-(2,3-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone.

m.p. 120° C. or more. ¹H NMR (300 MHz, CDCl₃): δ 7.17-7.53 (m, 3H), 7.61(dd, J=8.8, 2.3 Hz, 1H), 7.91-7.98 (m, 1H), 8.43 (d, J=2.3 Hz, 1H),11.80 (s, 1H).

<Example 58> Synthesis of2-(2,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 2.5-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 15 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 104.5 mg (85.0%) of thefinal desired compound,2-(2,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone.

m.p. 170-172° C. ¹H NMR (300 MHz, CDCl₃): δ 2.54 (s, 3H), 7.20-7.31 (m,3H), 7.51 (dd, J=7.6, 7.6 Hz, 1H), 7.73 (d, J=7.6 Hz, 1H), 8.21 (d,J=7.6 Hz, 1H), 14.19 (s, 1H).

<Example 59> Synthesis of2-(2,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 2.5-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 15 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 84.9 mg (69.0%) of thefinal desired compound,2-(2,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone.

m.p. 156-158° C. ¹H NMR (300 MHz, CDCl₃): δ 2.59 (s, 3H), 7.15-7.29 (m,3H), 7.43 (d, J=8.0 Hz, 2H), 8.12 (d, J=8.0 Hz, 1H), 14.21 (s, 1H).

<Example 60> Synthesis of2-(2,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 2.5-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 15 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 82.3 mg (66.9%) of thefinal desired compound,2-(2,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone.

m.p. 120° C. or more. ¹H NMR (300 MHz, CDCl₃): δ 2.57 (s, 3H), 7.15-7.30(m, 3H), 7.52 (d, J=8.6 Hz, 1H), 7.70 (dd, J=8.6, 1.9 Hz, 1H), 8.15 (s,1H), 14.17 (s, 1H).

<Example 61> Synthesis of2-(2,5-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-chlorobenzophosphorane compound (200 mg, 0.38 mmol),diisopropylethylamine (0.131 mL, 0.75 mmol, 2 eq), 2.5-difluorobenzoylchloride (0.071 mL, 0.57 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 15 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 37.6 mg (29.8%) of thefinal desired compound,2-(2,5-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone.

m.p. 147-150° C. ¹H NMR (300 MHz, CDCl₃): δ 7.26-7.43 (m, 3H), 7.69 (dd,J=8.6, 1.8 Hz, 1H), 7.75 (d, J=1.8 Hz, 1H), 8.41 (d, J=8.6 Hz, 1H),13.94 (s, 1H).

<Example 62> Synthesis of2-(2,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-chlorobenzophosphorane compound (200 mg, 0.38 mmol),diisopropylethylamine (0.131 mL, 0.75 mmol, 2 eq), 2.5-difluorobenzoylchloride (0.071 mL, 0.57 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 15 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 108.8 mg (86.4%) of thefinal desired compound,2-(2,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone.

m.p. 168-170° C. ¹H NMR (300 MHz, CDCl₃): δ 7.16-7.33 (m, 3H), 7.60 (d,J=8.9 Hz, 1H), 7.84 (dd, J=8.9, 2.5 Hz, 1H), 8.34 (d, J=2.5 Hz, 1H),13.77 (s, 1H).

<Example 63> Synthesis of2-(2,5-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-bromobenzophosphorane compound (200 mg, 0.35 mmol),diisopropylethylamine (0.121 mL, 0.70 mmol, 2 eq), 2,5-difluorobenzoylchloride (0.066 mL, 0.52 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 15 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 65.2 mg (49.4%) of thefinal desired compound,2-(2,5-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone.

m.p. 152-153° C. ¹H NMR (300 MHz, CDCl₃): δ 7.16-7.32 (m, 3H), 7.53 (d,J=8.9 Hz, 1H), 7.98 (dd, J=8.9, 2.3 Hz, 1H), 8.50 (d, J=2.3 Hz, 1H),13.77 (s, 1H).

<Example 64> Synthesis of2-(3,4-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 3.4-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 10 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 81 mg (66%) of the finaldesired compound,2-(3,4-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone.

m.p. 203-204° C. ¹H NMR (300 MHz, CDCl₃): δ 2.55 (s, 3H), 7.29-7.38 (m,1H), 7.45-7.58 (m, 3H), 7.73 (d, J=6.7 Hz, 1H), 8.20 (d, J=8.1 Hz, 1H),14.30 (s, 1H).

<Example 65> Synthesis of2-(3,4-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 3.4-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), 4-(dimethylamino)pyridine (5 mg)were stirred in methylene chloride (4 mL) for 2 hours, and stirred intoluene (9 mL) for 10 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 47 mg (38%) of the finaldesired compound,2-(3,4-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone.

m.p. 202-204° C. ¹H NMR (300 MHz, CDCl₃): δ 7.30-7.36 (m, 1H), 7.43 (d,J=6.9 Hz, 3H), 7.49-7.55 (m, 1H), 8.24 (d, J=8.7 Hz, 1H), 14.30 (s, 1H).

<Example 66> Synthesis of2-(3,4-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 3.4-difluorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 10 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 70.9 mg (57.6%) of thefinal desired compound,2-(3,4-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone.

m.p. 222-223° C. ¹H NMR (300 MHz, CDCl₃): δ 2.56 (s, 3H), 7.30-7.36 (m,1H), 7.42-7.47 (m, 1H), 7.49-7.56 (m, 2H), 7.70 (dd, J=8.5, 1.8 Hz, 1H),8.14 (s, 1H), 14.20 (s, 1H).

<Example 67> Synthesis of2-(3,4-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-chlorobenzophosphorane compound (200 mg, 0.38 mmol),diisopropylethylamine (0.131 mL, 0.75 mmol, 2 eq), 3,4-difluorobenzoylchloride (0.071 mL, 0.57 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 10 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 100.8 mg (80.0%) of thefinal desired compound,2-(3,4-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone.

m.p. 195-198° C. ¹H NMR (300 MHz, CDCl₃): δ 7.31-7.37 (m, 1H), 7.43-7.47(m, 1H), 7.50-7.61 (m, 2H), 7.83 (dd, J=8.9, 2.4 Hz, 1H), 8.32 (d, J=2.4Hz, 1H).

<Example 68> Synthesis of2-(4-chlorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 4-chlorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour, and stirred intoluene (9 mL) for 40 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 99 mg (81%) of the finaldesired compound,2-(4-chlorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone.

m.p. 217-219° C. ¹H NMR (300 MHz, CDCl₃): δ 2.53 (s, 3H), 7.46-7.53 (m,3H), 7.63 (dd, J=6.7, 2.0 Hz, 2H), 7.70 (d, J=6.7 Hz, 1H), 8.18 (d,J=8.0 Hz, 1H), 14.35 (s, 1H).

<Example 69> Synthesis of2-(4-chlorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-4-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 4-chlorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour, and stirred intoluene (9 mL) for 70 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 105 mg (85%) of thefinal desired compound,2-(4-chlorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone.

m.p. 220-222° C. ¹H NMR (300 MHz, CDCl₃): δ 2.56 (s, 3H), 7.40 (d, J=6.7Hz, 2H), 7.48 (dd, J=6.7, 2.0 Hz, 2H), 7.60 (dd, J=6.7, 2.0 Hz, 2H),8.22 (d, J=8.6 Hz, 1H), 14.35 (s, 1H).

<Example 70> Synthesis of2-(4-chlorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-methylbenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.136 mL, 0.80 mmol, 2 eq), 4-chlorobenzoylchloride (0.074 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour, and stirred intoluene (9 mL) for 60 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 57.0 mg (46.3%) of thefinal desired compound,2-(4-chlorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone.

m.p. 212-227° C. ¹H NMR (300 MHz, CDCl₃): δ 2.54 (s, 3H), 7.49 (dd,J=8.6, 2.7 Hz, 3H), 7.60 (dd, J=8.6, 1.8 Hz, 2H), 7.67 (dd, J=8.6, 2.1Hz, 1H), 8.12 (s, 1H), 14.50 (s, 1H).

<Example 71> Synthesis of2-(4-chlorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-chlorobenzophosphorane compound (200 mg, 0.38 mmol),diisopropylethylamine (0.131 mL, 0.75 mmol, 2 eq), 4-chlorobenzoylchloride (0.072 mL, 0.57 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour, and stirred intoluene (9 mL) for 80 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 48.2 mg (38.3%) of thefinal desired compound,2-(4-chlorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone.

m.p. 227-228° C. ¹H NMR (300 MHz, CDCl₃): δ 7.50 (d, J=8.5 Hz, 2H),7.55-7.62 (m, 3H), 7.80 (dd, J=8.9, 2.5 Hz, 1H), 8.30 (d, J=2.5 Hz, 1H),14.20 (s, 1H).

<Example 72> Synthesis of2-(4-chlorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-5-bromobenzophosphorane compound (200 mg, 0.35 mmol),diisopropylethylamine (0.121 mL, 0.70 mmol, 2 eq), 4-chlorobenzoylchloride (0.066 mL, 0.52 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 1 hour, and stirred intoluene (9 mL) for 40 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 61.4 mg (46.5%) of thefinal desired compound,2-(4-chlorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone.

m.p. 211-213° C. ¹H NMR (300 MHz, CDCl₃): δ 7.47-7.50 (m, 3H), 7.73 (d,J=8.5 Hz, 2H), 7.87 (dd, J=8.8, 2.3 Hz, 1H), 8.39 (d, J=2.3 Hz, 1H).

<Example 73> Synthesis of2-(4-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxybenzophosphorane compound (200 mg, 0.40 mmol),diisopropylethylamine (0.140 mL, 0.80 mmol, 2 eq), 4-chlorobenzoylchloride (0.077 mL, 0.60 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 17 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 47.1 mg (38.9%) of thefinal desired compound,2-(4-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone.

m.p. 212-213° C. ¹H NMR (300 MHz, CDCl₃): δ 7.50 (d, J=8.6 Hz, 2H),7.58-7.63 (m, 4H), 7.84-7.90 (m, 1H), 8.35 (dd, J=8.2, 1.7 Hz, 1H),14.35 (s, 1H).

<Example 74> Synthesis of2-(4-chlorophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone

Through the general synthesis method of Preparation Example 1 above, a2-hydroxy-3-methoxybenzophosphorane compound (200 mg, 0.38 mmol),diisopropylethylamine (0.132 mL, 0.76 mmol, 2 eq), 4-chlorobenzoylchloride (0.072 mL, 0.57 mmol, 1.5 eq), and 4-(dimethylamino)pyridine (5mg) were stirred in methylene chloride (4 mL) for 2 hours, and stirredin toluene (9 mL) for 20 hours, and then stirred in TFA (6 mL) andmethylene chloride (6 mL) for 2 hours to obtain 30.8 mg (24.4%) of thefinal desired compound,2-(4-chlorophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone.

m.p. 238-239° C. ¹H NMR (300 MHz, CDCl₃): δ 4.02 (s, 3H), 7.34 (d, J=8.0Hz, 1H), 7.49-7.54 (m, 3H), 7.67 (d, J=8.5 Hz, 2H), 7.88 (dd, J=8.0, 1.2Hz, 1H), 14.30 (s, 1H).

<Experimental Example 1> Measurement of TNF-Induced Cell DeathInhibitory Activity of the Synthesized 4-Benzopyranone DerivativeCompound

First, the addition of recombinant TNF to mouse fibroblast LM cellsinduces cell death, and thus, by using the same, the TNF-neutralizationbioassay was constructed. In other words, LM cells were pretreated withTNF (20 ng/ml) and the compounds synthesized in the above examples atvarious concentrations (5×10⁴ cells/well), and a sensitizer actinomycinD (0.5 μg/ml) was added, and cultured in a CO₂ incubator for 24 hours,and then the MTT assay was carried out. At this time, 10 μl of MTT (5mg/ml stock) was added, reacted for 4 hours, and then dissolved in DMSOto measure the absorbance at 570 nm. First, the TNF cell deathinhibition/neutralization action of TNF-target expected compounds atsingle concentration (50 or 10 μM) was measured. Second, IC50 valueswere determined by treating with the compounds having an excellenteffect at different concentrations. The % inhibition of TNF wascalculated using the following formula:

% inhibition of TNF=(OD value of a group treated with TNF andcompound−OD value of TNF treated group/(OD value of TNF untreatedgroup−OD value of TNF treated group)×100.

IC50 values were statistically analyzed using Prism 6 (GraphPad) (IC50:50% inhibitory concentration, CC50: 50% cytotoxic concentration).

As a result, as disclosed in Tables 1 and 2 below, the cytotoxicity ofTNF was inhibited up to 100% when treating with the compounds on thelist together in comparison with treating with TNF only.

TABLE 1 Concentration TNF inhibition IC50 CC50 Compound (μM) (%) (μM)(μM) Example 1 50 39.1 Example 2 50 46.6 Example 3 50 26.3 Example 4 5024.6 Example 5 50 34.4 Example 6 50 46.0 Example 7 50 47.0 Example 8 5024.3 Example 9 50 49.2 Example 10 50 45.6 Example 11 50 60.6 Example 1250 62.5 Example 13 50 55.9 Example 14 50 38.8 Example 15 50 47.5 Example16 50 70.2 16.8 >50 Example 17 50 71.0 2.5 >50 Example 18 50 48.6Example 19 50 48.3 Example 20 50 59.0 Example 21 50 55.2 Example 22 5050.4 Example 23 50 70.1 2.7 >50 Example 24 50 76.3 2.5 >50 Example 25 5041.0 Example 26 50 45.1 Example 27 50 65.2 Example 28 50 34.9 Example 2950 45.7 Example 30 50 7.4 Example 31 50 24.8 Example 32 50 23.6 Example33 50 20.7 Example 34 50 21.7 Example 35 50 28.1 Example 36 50 27.0Example 37 50 33.4

TABLE 2 IC50 CC50 Compound Concentration (μM) TNF inhibition (%) (μM)(μM) Example 38 50 30.9 Example 39 50 24.5 Example 40 50 100.0 122.4 >50Example 41 50 37.2 Example 42 50 23.0 Example 43 50 19.4 Example 44 5093.2 45.6 >50 Example 45 50 88.4 18.5 42.0 Example 46 50 100.0 28.9 >50Example 47 50 21.4 Example 48 50 29.0 Example 49 50 37.7 Example 50 50100.0 5.1 36.0 Example 51 50 100.0 4.5 13.0 Example 52 50 18.5 Example53 50 22.8 Example 54 50 26.5 Example 55 50 100.0 33.6 25.0 Example 5650 100.0 16.4 49.0 Example 57 50 100.0 10.3 47.0 Example 58 50 33.8Example 59 50 32.3 Example 60 50 28.1 Example 61 50 95.7 1.8 >50 Example62 50 100.0 3.8 17.0 Example 63 50 100.0 36.9 36.0 Example 64 50 47.2Example 65 50 25.1 Example 66 50 37.0 Example 67 50 100.0 3.9 21.0Example 68 50 15.8 Example 69 50 14.6 Example 70 50 28.9 Example 71 5097.9 3.3 36.0 Example 72 50 100.0 20.8 >50 Example 73 50 42.0 Example 7450 79.0 52.5 >50

Among these compounds, the compound of Example 61 had the lowest IC50value and cytotoxicity, so that the action mechanism and activity ofthis compound were studied more in depth. FIG. 1 shows theTNF-inhibiting levels of representative compounds including the compoundof Example 61 depending on concentration.

<Experimental Example 2> Measurement of TNF Direct Binding andTNF-Etanercept Binding Inhibitory Activity of 4-Benzopyranone DerivativeCompounds (1) Measurement of Real-Time Binding of 4-BenzopyranoneDerivative Compound to TNF Using SPR Analysis Method

The binding kinetics of TNF and 4-benzopyranone derivative compounds wasmeasured by Surface Plasmon Resonance (SPR) method. First, based on theresults of pH scouting (10 mM acetate buffer, pH 4.0, 4.5, 5.0, and5.5), the appropriate buffer pH was determined, and then the recombinantTNF was immobilized on the CM5 chip (4000-5000 RU). In order to correctan effect of DMSO, a solvent correction standard was then prepared, andDMSO was added to the HBS-EP+ buffer (GE) in an amount of 5%. Then, the4-benzopyranone derivative compounds were prepared depending onconcentration, flowed into the chip (20 μl/min, 500 s), and dissolved(500 s), and the results of sensorgram were analyzed by a dedicatedprogram to determine the affinity value (K_(D)). At this time, therunning buffer was HBS-EP+ buffer (GE), the regeneration buffer was 10mM glycine (pH 2.5), and Biacore T200 (GE) was used as an analyzer.

As a result, as shown in FIGS. 2 a to 2 e , the TNF direct binding ofthe compound of Example 61 was measured using SPR assay, and the bindingaffinity (K_(D)) was 1.439×10⁻⁶ (M) (FIGS. 2 a and 2 b ). It wasconfirmed that the compound of Example 61 did not bind to etanercept(FIG. 2 c ), and the compound of Example 30, which had almost no TNFneutralization effect (Table 1, 7.4% inhibition), did not bind to TNF(FIG. 2 d ).

(2) Measurement of TNF-Etanercept Binding Inhibitory Activity of4-Benzopyranone Derivative Compounds Using SPR Analysis Method

The competition of binding of TNF-inhibiting biopharmaceuticaletanercept and the derivative compounds to TNF was measured by SPRassay. Based on the results of pH scouting (10 mM acetate buffer, pH4.0, 4.5, 5.0, and 5.5), the appropriate buffer pH was determined, andthen the recombinant TNF was immobilized on the CM5 chip (4000-5000 RU).In order to correct an effect of DMSO, a solvent correction standard wasthen prepared, and DMSO was added to the PBST buffer (1×PBS, pH 7.4,0.05% Tween 20, 0.01% Triton X-100, 5% DMSO) in an amount of 5%. Then,Etanercept alone or in combination with the compounds was flowed intothe chip (20 μl/min, 300 s) and dissolved (700 s), and the results ofsensorgram were analyzed by a dedicated program. The changes in the RUvalue of the compound mixture samples were observed with comparison withthe biopharmaceuticals alone control group. At this time, the runningbuffer was PBST buffer, the regeneration buffer was 10 mM glycine (pH2.5), and the instrument was Biacore T200 (GE).

As a result, as shown in FIG. 2 e , the compound of Example 61significantly inhibited the TNF-etanercept binding. Thus, it was provedthat the compound and etanercept competed with each other against TNF,and the compound inhibited the binding of TNF to TNF receptor(etanercept) by binding to TNF.

<Experimental Example 3> Analysis of TNF-Cell Binding and TNF-CellSignaling Inhibitory Activity by 4-Benzopyranone Derivative Compounds(1) Measurement of Inhibitory Activity Against Binding to Raw 264.7 Cellof TNF by 4-Benzopyranone Derivative Compound Using Flow CytometryAnalysis Method

First, macrophage Raw 264.7 cells were prepared (4×10⁶ cells/ml), andthe biotinylated TNF alone or in combination with the 4-benzopyranonederivative compound (800, 200, 5 μM) was added to the cells and reacted(30 min, 4° C.). 10 μl of avidin-FITC reagent was then added and reactedin the dark (30 min, 4° C.). After washing with RDF1 buffer, FACSanalysis was carried out (BD Canto). Human TNF-alpha BiotinylatedFluorokine Flow Cytometry Kit (Cat #: NFTA0) of R&D Systems was used inthe experiment.

As a result, as shown in FIG. 3 a , the compound of Example 61 havingthe lowest IC50 value (Table 2, 1.8 μM) decreased the target cellbinding of TNF to the basal level in a concentration-dependent manner,and the compound of Example 51 having the relatively high IC50 value(Table 2, 4.5 μM) exhibited the moderate inhibitory activity against theTNF-cell binding, and the compound of Example 30 having almost noinhibitory activity against the TNF toxicity (Table 1) did not inhibitthe cell binding of TNF at all. Based on the above, it was found thatthere is a close correlation between the TNF cytotoxicity inhibitoryeffect and the TNF-cell binding inhibitory effect.

(2) Measurement of Inhibitory Activity Against LM Cell Signaling of TNFby 4-Benzopyranone Derivative Compounds Using Western Blot AnalysisMethod

First, LM cells was treated with TNF alone (50 ng/ml) or a mixture ofTNF and the compound of Example 61 (0, 10, 25, 50 μM) for 1 hour, andthen the cells were washed and prepared. After the cytoplasmic fractionand nuclear fraction were separated/obtained using NE-PER nuclear andcytoplasmic extraction reagents (Thermo Scientific, cat #78833), NF-kBp65 protein was detected by SDS-PAGE and Western Blot. Anti-NF-κB p65rabbit monoclonal antibody (Cell Signaling Technology, cat # D14E12) wasused as a primary antibody, and goat anti-rabbit HRP-conjugated antibody(Invitrogen, cat #656120) was used as a secondary antibody. As a WesternBlot control protein (housekeeping protein), GAPDH was used in thecytoplasmic fraction, and Lamin B1 was used in the nuclear fraction.Anti-GAPDH mouse monoclonal antibody (GenScript, cat # A01622) was usedas a primary antibody of GAPDH, and goat anti-mouse HRP-conjugatedantibody (BETHYLlaboratories, cat # A90-116P) was used as a secondaryantibody. Anti-Lamin-B1 rabbit monoclonal antibody (Cell SignalingTechnology, cat #12586S) was used as a primary antibody of Lamin B1, andanti-rabbit HRP-conjugated antibody (Invitrogen, cat #656120) was usedas a secondary antibody.

As a result, as shown in FIG. 3 b , it was found that the migration ofNF-kB from the cytoplasm to the nucleus activated by the TNF treatmentwas almost completely inhibited by the treatment of the compound ofExample 61 at about 25 μM.

Considering all the results of FIGS. 3 a and 3 b , the compound ofExample 61 also seems to inhibit the cell signaling of TNF byintrinsically blocking the cell binding of TNF.

<Experimental Example 4> Analysis of Therapeutic Effect on Sepsis by4-Benzopyranone Derivative Compounds (1) In Vivo TNF NeutralizationAssay (TNF-Induced Sepsis)

Treatment with TNF and a sensitizer D-galactosamine in an in vivo mousemodel leads to death from acute liver failure. Thus, the compound ofExample 61 (3.3 mpk or 16.5 mpk) was orally administered to BALB/c mice,and after 30 minutes, the mice were IP treated with the mixture ofD-galactosamine (21 mg/mouse) and TNF (0.3 μg/mouse). Survival rateswere observed and recorded every 3 hours up to 24 hours, andstatistically analyzed by log-rank (Mantel-Cox) test using Prism 6(GraphPad) (N=13/group, ** P<0.005, *** P<0.001). mpk, mg/kg). Thisconfirmed that the in vivo neutralization activity of the compoundsbinding to TNF in a TNF-induced lethality model.

As a result, as shown in FIG. 4 a , it was found that the death of micedue to acute hepatotoxicity induced by D-galactosamine and TNF wasinhibited by oral administration (PO) of the compound of Example 61(3.3, 16.5 mpk) in a concentration-dependent manner.

(2) Lipopolysaccharide (LPS)-Induced Sepsis Model

The sepsis mouse model was induced by the intraperitoneal infusion ofLPS (45 mg/kg; Sigma, E. coli 055:B5). The mouse survival experiment wascarried out by observing the survival rates every 24 hours up to 3 daysafter co-administering the compound of Example 61 (50 mg/kg) and LMT-28(50 mg/kg) immediately after the infusion of LPS.

As a result, as shown in FIG. 4 b, 100% survival rate was shown in thegroup treated with the compound of Example 61 and LMT-28 in comparisonwith the LPS alone treated group in which the survival rate was 20%after 72 hours (N=5/group, ***P<0.001).

<Experimental Example 5> Analysis of Prophylactic/Therapeutic Effect onRheumatoid Arthritis by 4-Benzopyranone Derivative Compounds (1)Prophylactic Effect on Rheumatoid Arthritis (TNF-Overexpressing MouseModel)

The TNF-overexpressing C57BL/6 mouse is a gene recombinant mouse, and amodel in which human TNF is overexpressed and arthritis is induced asthe age of the mouse becomes higher. The compound of Example 61 (3.3 mpkor 33 mpk) was orally administered to the TNF-overexpressing mice fromWeek 8 to Week 24, three times a week, and the pathology was measuredand recorded from Week 7 to Week 24. The pathology score was measured as1=erythema or bent foot, 2=bent foot and weak swelling, 3=bent foot andmoderate swelling, 4=ankle bending and weak swelling, 5=ankle bendingand severe swelling (N=4/group, * P<0.01, ** P<0.005, *** P<0.001).

As a result, as shown in FIG. 5 a , the TNF-inhibiting compoundsinhibited TNF-induced rheumatoid arthritis in a concentration-dependentmanner and showed a significant efficacy even at a low dose of 3.3 mpk.At the dose of 33 mpk, a remarkable prophylactic effect on rheumatoidarthritis was observed in comparison with the negative control group.

(2) Therapeutic Effect on Rheumatoid Arthritis (TNF-Overexpressing MouseModel)

First, until the mean level of arthritis of TNF-overexpressing micereached 8, the mice were raised. As such, etanercept (4.5 mpk),adalimumab (1.2 mpk), the compound of Example 61 (25 mpk, 50 mpk, or 100mpk) were orally administered to the mice with rheumatoid arthritis fromWeek 15 to Week 23, three times a week, and the pathology was measuredand recorded from Week 7 to Week 23 (N=5 to 6/group, * P<0.01, **P<0.005, *** P<0.001).

As a result, as shown in FIG. 5 b , the TNF-inhibiting compounds showeda therapeutic efficacy on rheumatoid arthritis already induced by TNF ina concentration-dependent manner. At all doses of 25, 50, and 100 mpk, aremarkable therapeutic effect on rheumatoid arthritis was observed incomparison with the negative control group. When the compound of Example61 was orally administered at the doses of 50 and 100 mpk, anon-inferior effect was observed even in comparison with adalimumab andetanercept, which are each commercially available injectablebiopharmaceuticals.

(3) Synergistic Therapeutic Effect on Rheumatoid Arthritis whenAdministered in Combination with Tofacitinib (CIA Mouse Model)

The CIA mouse model is a model in which arthritis is induced by collagenin DBA/1 mice. Thus, a 1:1 mixture of collagen type 2 and CFA wasintradermally (ID) injected into the base of the tail of 8-week oldDBA/1 mice. Two weeks after the primary immunization, a 1:1 mixture ofcollagen type 2 and IFA was intradermally (ID) injected into the base ofthe tail. After the second immunization, the pathology was measuredtwice a week, and the groups were divided equally so that the mean ofthe pathology of each group was 4 points. From Day 35 after the primaryimmunization, etanercept (4.5 mpk), adalimumab (1.2 mpk), the compoundof Example 61 (25 mpk, 50 mpk, or 100 mpk) were orally administeredthree times a week, and the pathology was measured and recorded. Thepathology score was measured as 1=inflammation and swelling of one toe,2=inflammation of one or more toes or weak swelling of the whole sole ofthe foot, 3=severe swelling of the whole sole of the foot and swellingof ankle, 4=very severe inflammation and swelling of the whole foot orthe bending and stiffness of foot (N=7 to 9/group, * P<0.01, ** P<0.005,*** P<0.001).

As a result, as shown in FIG. 5 c , the TNF-inhibiting compounds showedremarkable therapeutic efficacy on rheumatoid arthritis already inducedby the infusion of collagen. At all doses of 25, 50, and 100 mpk, aremarkable therapeutic effect on rheumatoid arthritis was observed incomparison with the negative control group. The compound of Example 61(25 mpk) showed considerably similar efficacy to tofacitinib (25 mpk),which is a commercially available oral JAK inhibitor, and showed anon-inferior effect even in comparison with etanercept, which is aninjectable biopharmaceuticals.

(4) Synergistic Therapeutic Effect on Rheumatoid Arthritis whenAdministered in Combination with Methotrexate (MTX) (CIA Mouse Model)

A 1:1 mixture of collagen type 2 and CFA was intradermally (ID) injectedinto the base of the tail of DBA/1 mice. Two weeks after the primaryimmunization, a 1:1 mixture of bovine collagen type 2 and IFA(incomplete Freund's adjuvant) was intradermally (ID) injected into thebase of the tail to prepare the rheumatoid arthritis animal model. Thethus prepared mice were divided into three groups, i.e., a positivecontrol group (MTX 5 mpk treated group: 10 mice), a group treated withthe compound of Example 61 alone (10 mpk treated group: 9 mice), a grouptreated with MTX and the compound of Example 61 (each 5 mpk and 10 mpk;9 mice), and a group treated with vehicle (a group treated with 0.05%CMC diluted in distilled water; 15 mice). At the time of the firstsymptom (Day 25 after the primary immunization), each of the drug wasintraperitoneally or orally administered three times a week (license#2016-018-02), and the compound of Example 61 was orally administeredonce a day.

The severity of arthritis was determined by ascertaining the mean scoreof arthritis according to the following standard score in the forefootand hindfoot twice a week from Day 0 to Day 73 after the primaryimmunization: 0=normal foot, 1=inflammation and swelling of one toe,2=inflammation and swelling of more than one toe, but not all of thetoes have inflammation and swelling, or weak swelling of all toes,3=inflammation and swelling of all toes, and 4=severe inflammation andswelling or occlusion of foot.

As a result, as shown in FIG. 8 , even when treating with only thecompound of Example 61 or MTX alone, the therapeutic effect on arthritiswas shown. However, when the two ingredients are used together, theremarkably excellent therapeutic effect on arthritis was shown. Thus, itwas confirmed that a remarkable synergistic effect was shown.

<Experimental Example 6> Analysis of Therapeutic Effect on InflammatoryBowel Disease (IBD) by 4-Benzopyranone Derivative Compounds

Inflammatory bowel disease was induced by the oral treatment withdextran sodium sulfate (DSS) to mice. The pathology was induced by DSS(2.5%, 6 days) in 7-week old C57BL/6 mice. At the same time, thecompound of Example 61 (100 mpk) and metformin (100 mpk) were orallyco-administered. After 9 days, the colon length of each treated groupwas measured, and the therapeutic effect of drugs on inflammatory boweldisease was statistically analyzed (N=5/group, * P<0.05, mean±SEM).

As a result, as shown in FIG. 6 , after the TNF inhibitory efficacy wasconfirmed in experiments in vitro/in vivo using the compound of Example61, the efficacy against inflammatory bowel disease was analyzed. As aresult, it was confirmed that the decrease of the colon length by DSSwas significantly inhibited when co-administering the compound ofExample 61 and metformin.

<Experimental Example 7> Analysis of Protective Effect on Sepsis-InducedAcute Kidney Injury by 4-Benzopyranone Derivative Compounds

The mice (male, C57/BL6, 12-14-week old, 25-30 g, N=5) were subjected tocecal ligation and puncture (CLP) or a sham procedure, and then werekept for 48 hours. The compound of Example 61 (50 mpk) was IP infusedimmediately after the CLP procedure. After 48 hours, in order to measurerenal function, the amount of blood urea nitrogen (BUN) and creatininein serum was analyzed.

As a result, as shown in FIG. 7 , the incidence of acute kidney injury(AKI) at 48 hours after the CLP procedure was about 75%, whereas in caseof the compound treated group, there was no significant increase orthere was a slight increase in the kidney injury marker in all of thetreated groups.

While the present invention has been particularly described withreference to specific embodiments thereof, it will be apparent to thoseskilled in the art that such specific embodiments are merely preferredembodiments and that the scope of the present invention is not limitedthereby. That is, the practical scope of the present invention isdefined by the appended claims and equivalents thereof

What is claimed is:
 1. A method for treating a TNF overexpressiondisease, comprising administering a pharmaceutically effective amount ofa composition to a subject in need thereof, wherein the compositioncomprises (i) a 4-benzopyranone derivative represented by Formula 1, apharmaceutically acceptable salt, solvate, racemate, or stereoisomerthereof:

in which, R¹ is heteroaryl or substituted phenyl; R² is hydrogen orlower alkyl of (C1-C4); and R³, R⁴, R⁵, and R⁶ are each independentlyhydrogen, halogen, or (C1-C4)alkoxy, and (ii) a drug selected from thegroup consisting of anti-rheumatic drugs (DMARDs), nonsteroidalanti-inflammatory drugs (NSAIDs), steroids, antimetabolites,anti-inflammatory agents, antibiotics, signaling/enzyme inhibitors,receptor inhibitors, HMGB1 inhibitors, antithrombotic drugs, autophagyagonists, cytokine inhibitors, HMG-CoA reductase inhibitors,antihypertensive agents, anticancer agents, immune activation agents, Bcell inhibitors, and T cell inhibitors.
 2. The method according to claim1, wherein in Formula 1, R¹ is furanyl, thiophenyl, or substitutedphenyl represented by

and R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ are each independentlyhydrogen, chloro, bromo, fluoro, or methoxy.
 3. The method for treatinga TNF overexpression disease, comprising administering apharmaceutically effective amount of a composition to a subject in needthereof, wherein the composition comprises a 4-benzopyranone derivative,pharmaceutically acceptable salt, solvate, racemate, or stereoisomerthereof, wherein the 4-benzopyranone derivative is selected from thegroup consisting of2-(2-chlorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2-furanyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3-methylphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-methoxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2-thiophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3,4-dimethoxyphenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-(4H)-4-benzopyranone; 2-(3-fluorophenyl)carboxy-8-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy chloro-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3-fluorophenyl)-3-carboxy-7-methoxy-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(3,5-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2,3-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(2,5-difluorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(3,4-difluorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-8-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-7-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-methyl-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-chloro-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-6-bromo-(4H)-4-benzopyranone;2-(4-chlorophenyl)-3-carboxy-(4H)-4-benzopyranone; and2-(4-chlorophenyl)-3-carboxy-8-methoxy-(4H)-4-benzopyranone.
 4. Themethod according to claim 1, wherein the TNF-related disease is any oneselected from the group consisting of autoimmune diseases, inflammatorydiseases, cardiovascular diseases, metabolic diseases, immune disorders,neurological diseases, ophthalmic diseases, skin diseases, psychiatricdiseases, infectious diseases, and cancers.
 5. The method according toclaim 1, wherein the TNF-related disease is selected from the groupconsisting of rheumatoid arthritis, juvenile rheumatoid arthritis,inflammatory bowel disease, Crohn's disease, ulcerative colitis,psoriasis, plaque psoriasis, juvenile plaque psoriasis, psoriaticarthritis, polyarticular juvenile idiopathic arthritis, Behcet'senteritis, ankylosing spondylitis, axial spondyloarthritis, juvenileenthesitis-related arthritis, polymyalgia rheumatica, multiplesclerosis, thyroiditis, delayed hypersensitivity, allergy, contactdermatitis, atopic dermatitis, systemic lupus erythematosus, systemicsclerosis, adult-onset Still's disease, asthma, autoimmune thyroiddisorder, Sjogren's syndrome, Kawasaki disease, pancreatitis, nephritis,hepatitis, pneumonia, chronic obstructive pulmonary disease, otitismedia, angioplasia nephritis, myelodysplastic syndrome, osteoarthritis,sarcoidosis, granuloma annulare, Wegener's granulomatosis, lupus,hemolytic uremic syndrome, arteriosclerosis, vasculitis, heart failure,stroke, myocardial infarction, myocardial ischemia-reperfusion injury,sexual dysfunction, obesity, hypertension, diabetes mellitus anddiabetic complication, hyperlipidemia, preeclampsia, kidney disease,liver disease, kidney injury, liver injury, snake bite, allograftrejection, organ transplantation, graft versus host disease, dementia,Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis,pain, central nervous system disease, uveitis, Behcet's disease,diabetic macular edema, macular degeneration, orbitopathy, glaucoma,hidradenitis suppurativa, multicentric reticulohistiocytosis, pityriasisrubra pilaris, eosinophilic fascilitis, panniculitis, necrobiosislipoidica diabeticorum, cicatricial pemphigoid, pyoderma gangrenosum,Sweet's syndrome, subcorneal pustular dermatosis, scleroderma,neutrophilic dermatitis, toxic epidermal necrolysis, pustulardermatitis, dermatomyositis, polymyositis, bullous dermatosis, erythemanodosum, alopecia, depressive disorder, bipolar disorder, anxietydisorder, tuberculosis, viral infection, bacterial infection, fungalinfection, protozoan infection, cerebral malaria, sepsis, septic shock,prostate cancer, skin cancer, colorectal cancer, kidney cancer,pancreatic cancer, ovarian cancer, breast cancer, bladder cancer,prostate cancer, lymphoma, glioma, osteosarcoma, leukemia, multiplemyeloma, and cachexia.
 6. The method according to claim 1, wherein thedrug is selected from the group consisting of methotrexate,hydroxychloroquine, sulfasalazine, leflunomide, bucillamine,cyclosporine, tacrolimus, azathioprine, cyclophosphamide, mizoribine,penicillamine, oral gold preparation, antimalarial agent,6-mercaptopurine, indomethacin, naproxen, sulindac, diclofenac,aceclofenac, mefenamic acid, aspirin, fenoprofen, salsalate, piroxicam,etodolac, flurbiprofen, ibuprofen, loxoprofen, nabumetone, lonazolac,meloxicam, fenbufen, ketorolac tromethamine, indoprofen, ketoprofen,suprofen, carprofen, tiaprofenic acid, flufenamic acid, ebselen,felbinac, tolmetin, flunixin, celecoxib, rofecoxib, hydrocortisone,cortisone, prednisolone, methylprednisolone, triamcinolone,betamethasone, dexamethasone, fludrocortisone, entocort,5-aminosalicylate, 6-thioguanine, cytarabine, 5-fluorouracil,dacarbazine, mechlorethamine, thiotepa, chlorambucil, melphalan,carmustine, lomustine, busulfan, sestrin 2, withaferin A, celastrol,quercetin, luteolin, curcumin, metformin, dibromomannitol, GR270773,pentoxifylline, N-acetylcysteine, melatonin, resveratrol, mesalamine,single chain fatty acid, glutamine, gemfibrozil, retinoid, hydroxyurea,trihydroxyisoflavone, deoxykaempferol, kaempferol, gingerol, caffeicacid, cyanidin, cryptotanshinone, deguelin, delphinidin, equol, fisetin,myricetin, procyanidin B2, metronidazole, ciprofloxacin, niclosamide,thiabendazole, imipenem-cilastatin, fluoroquinolone, tofacitinib,glyburide, rolipram, doxycycline, VX-166, zVAD, L-97-1, ISO-1,tauroursodeoxycholic acid, HK-156, A-285222, CP-0127,Bis-N-norgliovictin, aurintricarboxylic acid, chloroamidine, ouabain,terazosin, prazosin, tranilast, apremilast, monobenzone,phenazopyridine, 546C88, NOX-100, gabexate mesilate, ulinastatin,somatostatin, octreotide, IKK inhibitor, caspase inhibitor, TAK-242,eritoran, ki16425, camptothecin, caffeic acid phenethyl ester,sulforaphane, Tim-3, BN-52021, BB-882, TCV-309, CT-400, ethyl pyruvate,hemin, CORM-2, tanshinone IIA sulfonate, nicotine, EGCG,isorhamnetin-3-O-galactoside, persicarin, catechin, carbenoxolone,glycyrrhizin, emodin-6-O-b-D-glucoside, acteoside, forsythoside B,rosmarinic acid, chlorogenic acid, inflachromene, cilostazol,clopidogrel, sarpogrelate, drotrecogin alpha, carbamazepine,chloroquine, anakinra, tocilizumab, LMT-28,1-(3-dimethylaminopropyl)-3-ethylurea, gp13 OF c, beta-arrestin 2,IL-30, diacerein, secukinumab, ustekinumab, ixekizumab, thalidomide,adalimumab, infliximab, pravastatin, atorvastatin, rosuvastatin,simvastatin, losartan, telmisartan, hydrochlorothiazide, furosemide,propranolol, metoprolol, captopril, amlodipine, clonidine, methyldopa,minoxidil, streptozotocin, mitomycin, cisplatin, daunorubicin,doxorubicin, dactinomycin, bleomycin, mithramycin, anthramycin,calicheamicin, duocarmycin, vincristine, taxol, docetaxel, cytochalasinB, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide,teniposide vinblastine, colchicine, dihydroxy anthracenedione,mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone,procaine, tetracaine, lidocaine, propranolol, tamoxifen, bazedoxifene,puromycin, anetholtrithion, nivolumab, pembrolizumab, ipilimumab,atezolizumab, alpha-galactosylceramide, SRT3025, DTA-1, IL-7, IL-2,IL-15, CXCL1, ATRA, gemcitabine, carboplatin, NCX-4016, CDDO-Me,sunitinib, zoledronic acid, Astragalus polysaccharide, rituximab,imuran, abatacept, GW9662, rosiglitazone, Y-27632, and alefacept.